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Sample records for dolichos lablab lectin

  1. Isolation and properties of a lectin from the seeds of the Indian bean or lablab (Dolichos lablab L.).

    PubMed

    Güran, A; Tichá, M; Filka, K; Kocourek, J

    1983-03-01

    The lectin of the Indian bean or lablab (Dolichos lablab L.) was purified by affinity chromatography on two types of affinity carriers: O-alpha-D-mannopyranosyl-Separon and Separon-bound ovomucoid. The lectin is homogeneous in the ultracentrifuge: S20, w = 6.14 S, Mr = 110 000; the molecule appears to comprise two pairs of two types of subunits (Mr 16 000 and 40 000), and contains 2% neutral sugar and 0.2 Mn and 0.5 Zn atom respectively. The lectin agglutinates human erythrocytes non-specifically with regard to ABO grouping at a limit concentration of 8 micrograms/ml, and this activity is inhibited most effectively by N-acetyl-D-glucosamine, methyl alpha-D-mannopyranoside and ovomucoid, but not by free D-mannose.

  2. N-glycan analysis of mannose/glucose specific lectin from Dolichos lablab seeds.

    PubMed

    B S, Gnanesh Kumar; Pohlentz, Gottfried; Schulte, Mona; Mormann, Michael; Nadimpalli, Siva Kumar

    2014-08-01

    An affinity purified mannose/glucose specific lectin from the seeds of Dolichos lablab (Indian bean/lablab bean) resolves into five subunits upon SDS-PAGE in the range of Mr 12-20kDa. Partial de novo sequencing of subunits resulted in 88% and 73% sequence coverage for α and β subunits of the cDNA derived FRIL (Flt3 receptor interacting lectin) sequence, respectively and suggested that four bands correspond to the α-subunits while the band of lowest molecular mass is designated as β. It was proposed in an earlier study on FRIL that the difference in molecular mass of α-subunits is due to differences in C-terminal processing and differential N-glycosylation i.e. numbers of N-glycans present (Colucci et al., 1999). Thus, differential N-glycosylation of the purified mannose/glucose specific lectin was unravelled by in-gel trypsin/chymotrypsin digestion of the α-subunits followed by desalting and ZIC-HILIC enrichment of N-glycopeptides. Subsequently, analyses by nano electrospray ionisation quadrupole time of flight mass spectrometry and low-energy collision-induced dissociation experiments revealed the presence of a typical paucimannose type N-glycan (Man2(Xyl)GlcNAc2(Fuc)) in α subunits 2-4.

  3. Affinity of a galactose-specific legume lectin from Dolichos lablab to adenine revealed by X-ray cystallography.

    PubMed

    Shetty, Kartika N; Latha, Vakada Lavanya; Rao, Rameshwaram Nagender; Nadimpalli, Siva Kumar; Suguna, Kaza

    2013-07-01

    Crystal structure analysis of a galactose-specific lectin from a leguminous food crop Dolichos lablab (Indian lablab beans) has been carried out to obtain insights into its quaternary association and lectin-carbohydrate interactions. The analysis led to the identification of adenine binding sites at the dimeric interfaces of the heterotetrameric lectin. Structural details of similar adenine binding were reported in only one legume lectin, Dolichos biflorus, before this study. Here, we present the structure of the galactose-binding D. lablab lectin at different pH values in the native form and in complex with galactose and adenine. This first structure report on this lectin also provides a high resolution atomic view of legume lectin-adenine interactions. The tetramer has two canonical and two DB58-like interfaces. The binding of adenine, a non-carbohydrate ligand, is found to occur at four hydrophobic sites at the core of the tetramer at the DB58-like dimeric interfaces and does not interfere with the carbohydrate-binding site. To support the crystallographic observations, the adenine binding was further quantified by carrying out isothermal calorimetric titration. By this method, we not only estimated the affinity of the lectin to adenine but also showed that adenine binds with negative cooperativity in solution.

  4. Crystallization and preliminary X-ray crystallographic analysis of a galactose-specific lectin from Dolichos lablab

    SciTech Connect

    Lavanya Latha, V.; Kulkarni, Kiran A.; Nagender Rao, R.; Siva Kumar, N.; Suguna, K.

    2006-02-01

    The galactose-specific lectin from the seeds of a leguminous plant, D. lablab, has been crystallized. Molecular-replacement solution using 3.0 Å X-ray diffraction data showed the lectin to be a tetramer. The galactose-specific lectin from the seeds of Dolichos lablab has been crystallized using the hanging-drop vapour-diffusion technique. The crystals belong to space group P1, with unit-cell parameters a = 73.99, b = 84.13, c = 93.15 Å, α = 89.92, β = 76.01, γ = 76.99°. X-ray diffraction data to a resolution of 3.0 Å have been collected under cryoconditions (100 K) using a MAR imaging-plate detector system mounted on a rotating-anode X-ray generator. Molecular-replacement calculations carried out using the available structures of legume lectins as search models revealed that the galactose-specific lectin from D. lablab forms a tetramer similar to soybean agglutinin; two such tetramers are present in the asymmetric unit.

  5. Immunomodulatory Glc/Man-directed Dolichos lablab Lectin (DLL) evokes anti-tumor response in-vivo by counteracting angiogenic gene expressions.

    PubMed

    Vigneshwaran, V; Thirusangu, Prabhu; Br, Vijay Avin; Krishna, V; Pramod, Siddanakoppalu N; Prabhakar, B T

    2017-03-07

    Neovascularization and jeopardized immunity has been critically emphasized for the establishment of malignant progression. Lectins are the diverse class of carbohydrate interacting proteins having great potential as immunopotentiating and anticancer agents. The present investigation sought to demonstrate the antiproliferative activity of Dolichos lablab lectin (DLL) encompassing immunomodulatory attribute. DLL specific to glucose and mannose carbohydrate moieties has been purified to homogeneity from the common dietary legume Dolichos lablab. Results elucidated that DLL nonspecifically agglutinated blood cells and displayed striking mitogenecity to human and murine lymphocytes in-vitro with IL-2 production. The DLL conditioned medium exerted cytotoxicity towards malignant cells and neoangiogenesis in-vitro. Similarly, in-vivo antitumor investigation of DLL elucidated the regressed proliferation of ascitic and solid tumor cells which was paralleled with blockade of tumor neovasculature. DLL treated mice showed an upregulated immunoregulatory cytokine IL-2 in contrast to severely declined levels in control mice. Mechanistic validation revealed that DLL has abrogated the microvessel formation by weakening the proangiogenic signals specifically NF-κB, HIF-1 α, MMP-2&9 and VEGF in malignant cells leading to tumor regression. In summary, it is evident that the dietary lectin DLL potentially dampens the malignant establishment by mitigating neo-angiogenesis and immune shutdown. This study for the first time dictates the critical role of DLL as an immunostimulatory and anti-angiogenic molecule in cancer therapeutics. This article is protected by copyright. All rights reserved.

  6. Tryptophan environment, secondary structure and thermal unfolding of the galactose-specific seed lectin from Dolichos lablab: fluorescence and circular dichroism spectroscopic studies.

    PubMed

    Sultan, Nabil Ali Mohammed; Rao, Rameshwaram Nagender; Nadimpalli, Siva Kumar; Swamy, Musti J

    2006-07-01

    Fluorescence and circular dichroism spectroscopic studies were carried out on the galactose-specific lectin from Dolichos lablab seeds (DLL-II). The microenvironment of the tryptophan residues in the lectin under native and denaturing conditions were investigated by quenching of the intrinsic fluorescence of the protein by a neutral quencher (acrylamide), an anionic quencher (iodide ion) and a cationic quencher (cesium ion). The results obtained indicate that the tryptophan residues of DLL-II are largely buried in the hydrophobic core of the protein matrix, with positively charged side chains residing close to at least some of the tryptophan residues under the experimental conditions. Analysis of the far UV CD spectrum of DLL-II revealed that the secondary structure of the lectin consists of 57% alpha-helix, 21% beta-sheet, 7% beta-turns and 15% unordered structures. Carbohydrate binding did not significantly alter the secondary and tertiary structures of the lectin. Thermal unfolding of DLL-II, investigated by monitoring CD signals, showed a sharp transition around 75 degrees C both in the far UV region (205 nm) and the near UV region (289 nm), which shifted to ca. 77-78 degrees C in the presence of 0.1 M methyl-beta-D-galactopyranoside, indicating that ligand binding leads to a moderate stabilization of the lectin structure.

  7. Penetration of Crotalaria juncea, Dolichos lablab, and Sesamum indicum Roots by Meloidogyne javanica

    PubMed Central

    Araya, M.; Caswell-Chen, E. P.

    1994-01-01

    Penetration of Crotalaria juncea (PI 207657 and cv. Tropic Sun) Dolichos lablab cv. Highworth, and Sesamum indicum by juveniles (J2) of Meloidogyne javanica was assessed to investigate the mechanism by which these plants may reduce nematode numbers in the field. Growth chamber experiments were conducted at 25 C, with vials containing 90 g sand infested with 450 J2; tomato (UC 204 C) was included as a susceptible host. Fifteen days after inoculation, roots were stained and the nematodes within stained roots were counted. Both C. juncea lines were highly resistant to penetration, as they contained significantly fewer nematodes per cm of root and per root system than the other plants. Although containing more nematodes per cm of root than C. juncea, S. indicum and D. lablab had significantly fewer nematodes per root system and per cm of root than tomato. Roots were significantly longer in the plants with the lowest nematode penetration. Although C. juncea, D. lablab, and S. indicum may have potential utility as cover or rotation crops in soil infested with M. javanica, further quantitative information on the reproduction of M. javanica and other nematodes in these plants is needed. PMID:19279887

  8. In vivo biosynthetic studies of the Dolichos biflorus seed lectin

    SciTech Connect

    Quinn, J.M.; Etzler, M.E. )

    1989-12-01

    The in vivo biosynthesis of the Dolichos biflorus seed lectin was studied by pulse-chase labeling experiments using ({sup 35}S)methionine and ({sup 14}C)glucosamine. These studies demonstrate that each of the two mature lectin subunit types are derived by the processing of separate glycosylated precursors. The appearance of the precursor to subunit I before the precursor to subunit II supports the possibility raised by previous studies that both subunit types of this lectin may originate from a single gene product.

  9. Purification and characterization of a polyphenol oxidase from the seeds of field bean (Dolichos lablab).

    PubMed

    Paul, B; Gowda, L R

    2000-09-01

    The polyphenol oxidase from field bean (Dolichos lablab) seeds has been purified to apparent homogeneity by a combination of ammonium sulfate precipitation, DEAE-Sephacel chromatography, phenyl agarose chromatography, and Sephadex G-200 gel filtration. The purified enzyme has a molecular weight of 120 +/- 3 kDa and is a tetramer of 30 +/- 1.5 kDa. Native polyacrylamide gel electrophoresis of the purified enzyme revealed the presence of a single isoform with an observed pH optimum of 4.0. 4-Methyl catechol is the best substrate, followed by catechol, and L-3,4-dihydroxyphenylalanine, all of which exhibited a phenomenon of inhibition by excess substrate. No activity was detected toward chlorogenic acid, catechin, caffeic acid, gallic acid, and monophenols. Tropolone, both a substrate analogue and metal chelator, proved to be the most effective competitive inhibitor with an apparent K(i) of 5.8 x 10(-)(7) M. Ascorbic acid, metabisulfite, and cysteine were also competitive inhibitors.

  10. Purification and characterization of a proteinase inhibitor from field bean, Dolichos lablab perpureus L.

    PubMed

    Devaraj, V R; Manjunatha, N H

    1999-01-01

    A proteinase inhibitor resembling Bowman-Birk family inhibitors has been purified from the seeds of cultivar HA-3 of Dolichos lablab perpureus L. The protein was apparently homogeneous as judged by SDS-PAGE, PAGE, IEF, and immunodiffusion. The inhibitor had 12 mole% 1/2-cystine and a few aromatic amino acids, and lacks tryptophan. Field bean proteinase inhibitor (FBPI) exhibited a pI of 4.3 and an Mr of 18,500 Da. CD spectral studies showed random coiled secondary structure. Conformational changes were detected in the FBPI-trypsin/chymotrypsin complexes by difference spectral studies. Apparent Ka values of complexes of inhibitor with trypsin and chymotrypsin were 2.1x10(7) M(-1) and 3.1x10(7) M(-1), respectively. The binary and ternary complexes of FBPI with trypsin and chymotrypsin have been isolated indicating 1:1 stoichiometry with independent sites for cognate enzymes. Amino acid modification studies showed lysine and tyrosine at the reactive sites of FBPI for trypsin and chymotrypsin, respectively.

  11. Diphenol activation of the monophenolase and diphenolase activities of field bean (Dolichos lablab) polyphenol oxidase.

    PubMed

    Gowda, Lalitha R; Paul, Beena

    2002-03-13

    This paper reports a study on the hydroxylation of ferulic acid and tyrosine by field bean (Dolichos lablab) polyphenol oxidase, a reaction that does not take place without the addition of catechol. A lag period similar to the characteristic lag of tyrosinase activity was observed, the length of which decreased with increasing catechol concentration and increased with increasing ferulic acid concentration. The activation constant K(a) of catechol for ferulic acid hydroxylation reaction was 5 mM. The kinetic parameters of field bean polyphenol oxidase toward ferulic acid and tyrosine were evaluated in the presence of catechol. 4-Methyl catechol, L-dihydroxyphenylalanine, pyrogallol, and 2,3,4-trihydroxybenzoic acid, substrates with high binding affinity to field bean polyphenol oxidase, could stimulate this hydroxylation reaction. In contrast, diphenols such as protocatechuic acid, gallic acid, chlorogenic acid, and caffeic acid, which were not substrates for the oxidation reaction, were unable to bring about this activation. It is most likely that only o-diphenols that are substrates for the diphenolase serve as cosubstrates by donating electrons at the active site for the monophenolase activity. The reaction mechanism for this activation is consistent with that proposed for tyrosinase (Sanchez-Ferrer, A.; Rodriguez-Lopez, J. N.; Garcia-Canovas, F.; Garcia-Carmona, F. Biochim. Biophys. Acta 1995, 1247, 1-11). The presence of o-diphenols, viz. catechol, L-dihydroxyphenylalanine, and 4-methyl catechol, is also necessary for the oxidation of the diphenols, caffeic acid, and catechin to their quinones by the field bean polyphenol oxidase. This oxidation reaction occurs immediately with no lag period and does not occur without the addition of diphenol. The kinetic parameters for caffeic acid (K(m) = 0.08 mM, V(max) = 32440 u/mg) in the presence of catechol and the activation constant K(a) of catechol (4.6 mM) for this reaction were enumerated. The absence of a lag

  12. The conformational state of polyphenol oxidase from field bean (Dolichos lablab) upon SDS and acid-pH activation.

    PubMed

    Kanade, Santosh R; Paul, Beena; Rao, A G Appu; Gowda, Lalitha R

    2006-05-01

    Field bean (Dolichos lablab) contains a single isoform of PPO (polyphenol oxidase)--a type III copper protein that catalyses the o-hydroxylation of monophenols and oxidation of o-diphenols using molecular oxygen--and is a homotetramer with a molecular mass of 120 kDa. The enzyme is activated manyfold either in the presence of the anionic detergent SDS below its critical micellar concentration or on exposure to acid-pH. The enhancement of kcat upon activation is accompanied by a marked shift in the pH optimum for the oxidation of t-butyl catechol from 4.5 to 6.0, an increased sensitivity to tropolone, altered susceptibility to proteolytic degradation and decreased thermostability. The Stokes radius of the native enzyme is found to increase from 49.1+/-2 to 75.9+/-0.6 A (1 A=0.1 nm). The activation by SDS and acid-pH results in a localized conformational change that is anchored around the catalytic site of PPO that alters the microenvironment of an essential glutamic residue. Chemical modification of field bean and sweet potato PPO with 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide followed by kinetic analysis leads to the conclusion that both the enzymes possess a core carboxylate essential to activity. This enhanced catalytic efficiency of PPO, considered as an inducible defence oxidative enzyme, is vital to the physiological defence strategy adapted by plants to insect herbivory and pathogen attack.

  13. Effect of electroplating factory effluent on the germination and growth of hyacinth bean and mustard. [Dolichos lablab; Brassica compestris

    SciTech Connect

    Ajmal, M.; Khan, A.U.

    1985-12-01

    The effect of electroplating factory effluent in different concentrations (viz., 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, and 4.0%) on the germination and growth of hyacinth beans (Dolichos lablab) and mustard seeds (Brassica compestris) was studied. The germination of seeds was delayed with the increase of effluent concentration and the germination of mustard seeds was totally inhibited at 1.5% effluent concentration while hyacinth bean seeds tolerated the effluent up to 2.5% concentration. The metal content in the hyacinth bean plants increased with increasing effluent concentration but after 1.0% effluent concentration, the concentration of all the metals (Ca, Mg, Na, K, Cu, Zn, Fe) decreased in the plants except Cr, which increased throughout. Percentage germination, fresh weight, dry weight, root length, and shoot length of the plants were also analyzed. Cd, Ni, Co, Mn, and Pb were not detectable in the hyacinth bean plants.

  14. Development and Distribution of Dolichos biflorus Lectin as Measured by Radioimmunoassay 1

    PubMed Central

    Talbot, Craig F.; Etzler, Marilynn E.

    1978-01-01

    A radioimmunoassay, capable of detecting the Dolichos biflorus lectin at concentrations as low as 400 ng/ml, was developed and used to follow the distribution of this lectin in the plant during its life cycle. The lectin was first detected in the seeds of the plant 27 days after flowering and rapidly attained the high level of lectin present in the mature seed. The lectin content of the plant is highest in the seeds and cotyledons and decreases as the storage materials of the cotyledons decrease. A low but measurable amount of material that reacts with antibodies to the seed lectin was detected in the leaves, stems, and pods of the plant. This material gives a precipitin band of only partial identity to the seed lectin when tested in immunodiffusion against antiserum to the seed lectin. No lectin was detected by the radioimmunoassay in the roots of the plant at any stage of development. ImagesFIG. 4 PMID:16660399

  15. Crystallization of two related lectins from the legume plant Dolichos biflorus.

    PubMed

    Dao-Thi, M H; Hamelryck, T W; Bouckaert, J; Körber, F; Burkow, V; Poortmans, F; Etzler, M; Strecker, G; Wyns, L; Loris, R

    1998-11-01

    The seed lectin DBL and the related stem and leaves lectin DB58 of the tropical legume Dolichos biflorus were crystallized, as well as complexes of DBL with adenine and with GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal. The different crystal forms of DBL diffract to about 2.8 A, while DB58 crystals diffract to 3.3 A.

  16. Supplemental UV-B radiation induced changes in growth, pigments and antioxidant pool of bean (Dolichos lablab) under field conditions.

    PubMed

    Singh, Raghubansh; Singh, Suruchi; Tripathi, Ruchika; Agrawal, S B

    2011-03-01

    Present study is conducted to evaluate the response of bean (Dolichos lablabcv . pusa early prolific) plants to supplemental UV-B (sUV-B: 280-315 nm: 7.2 kJ m(-2) d(-1)) radiation. UV-B caused alteration in biomass translocation pattern with more retention of biomass in below ground parts leading to an increment in root shoot ratio. Specific leaf area (SLA) which is the measure of leaf thinness, increased in plants under sUV-B exposure by 95.7 and 82.3% after 15 and 30 days after germination. Photosynthetic machinery of bean plants was the potential target of UV-B as photosynthetic rate was decreased by 88.6% at 30 days after germination. sUV-B lead to the formation of reactive oxygen species thus generating oxidative stress. Stimulation of antioxidant defense system (enzymatic and non-enzymatic) was observed due to sUV-B radiation. Phenolic content decreased (34.7 and 18.6%) but protein showed varied response, increased initially (34%) thereafter declined (10.2%) under sUV-B radiation.

  17. Purification of alpha-mannosidase activity from Indian lablab beans.

    PubMed

    Tulasi, R B; Nadimpalli, S K

    1997-04-01

    Seeds of Dolichos lablab var. typicus (Indian lablab beans) contain a glucose/ mannose specific lectin that was affinity purified on Sepharose mannose columns in our laboratory. The unbound fraction from this matrix showed alpha-mannosidase activity. In the present study this has been purified to homogeneity by a combination of ion-exchange, hydrophobic chromatography and gel filtration. Purified alpha-mannosidase had an apparent molecular weight of 195,000 +/- 5,000 with 4.5% carbohydrate. On SDS-PAGE under reducing conditions, the enzyme dissociated into two major bands corresponding to Mr 66,000 and Mr 44,000. An antibody to the well studied jack bean alpha-mannosidase cross-reacts with the enzyme from the lablab beans suggesting antigenic similarity between these two legume mannosidases.

  18. Dual-function protein in plant defence: seed lectin from Dolichos biflorus (horse gram) exhibits lipoxygenase activity

    PubMed Central

    Roopashree, Subbaiah; Singh, Sridevi Annapurna; Gowda, Lalitha R.; Rao, A. G. Appu

    2006-01-01

    Plant–pathogen interactions play a vital role in developing resistance to pests. Dolichos biflorus (horse gram), a leguminous pulse crop of the subtropics, exhibits amazing defence against attack by pests/pathogens. Investigations to locate the possible source of the indomitable pest resistance of D. biflorus, which is the richest source of LOX (lipoxygenase) activity, have led to a molecule that exhibits LOX-like functions. The LOX-like activity associated with the molecule, identified by its structure and stability to be a tetrameric lectin, was found to be unusual. The evidence for the lectin protein with LOX activity has come from (i) MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS, (ii) N-terminal sequencing, (iii) partial sequencing of the tryptic fragments of the protein, (iv) amino acid composition, and (v) the presence of an Mn2+ ion. A hydrophobic binding site of the tetrameric lectin, along with the presence of an Mn2+ ion, accounts for the observed LOX like activity. This is the first ever report of a protein exhibiting both haemagglutination and LOX-like activity. The two activities are associated with separate loci on the same protein. LOX activity associated with this molecule adds a new dimension to our understanding of lectin functions. This observation has wide implications for the understanding of plant defence mechanisms against pests and the cellular complexity in plant–pathogen interactions that may lead to the design of transgenics with potential to impart pest resistance to other crops. PMID:16441240

  19. The lectin of Dolichos biflorus agglutinin recognises glycan epitopes on the surface of a subset of cardiac progenitor cells.

    PubMed

    Chen, Zhanfeng; Wang, Man; Xiang, Qiang; Sun, Zhenliang; Zhang, Rong

    2013-11-01

    The discovery of adult cardiac progenitor cells (CPCs) provides a promising way for treating heart disease; however, their surface characteristics that play a critical role in regulating their maintenance, self-renewal, migration, and differentiation have not been fully investigated. One subpopulation of Dolichos biflorus agglutinin (DBA)-positive cells was identified in the heart of adult mice. Flow cytometry showed that 3.7% of heart cells could be labeled by FITC conjugated DBA. BrdU pulse-chase showed that 55-75% of DBA(+) cells were CPCs. Evidences from 5-FU-induced myelosuppression along with BrdU pulse-chasing suggests that DBA-positive cells are proliferative. Furthermore, DBA positive cells display a cologenic appearance in vivo. Our findings suggest that DBA-positive cells in the heart of adult mouse contained a subset of CPCs, and DBA reactivity is one novel surface characteristic on CPCs.

  20. Lectin binding patterns in normal and neoplastic colonic mucosa. A study of Dolichos biflorus agglutinin, peanut agglutinin, and wheat germ agglutinin.

    PubMed

    Campo, E; Condom, E; Palacín, A; Quesada, E; Cardesa, A

    1988-11-01

    The cellular distribution of the carbohydrates labeled by Dolichos bifluorus agglutinin (DBA), peanut agglutinin (PNA), and wheat germ agglutinin (WGA) were studied in 21 normal colonic mucosae, 17 transitional mucosae, 9 nonneoplastic polyps (NNP), 27 adenomas, and 25 colorectal carcinomas. In normal mucosa DBA bound selectively to mucin of the goblet cells in the upper colonic crypt and to apical cytoplasm of the superficial columnar cells with a strong linear pattern. PNA binding was present only in the supranuclear portion (Golgi area) of the cells. WGA showed a strong reactivity in the goblet-cell mucin and in the supranuclear portion and apical cytoplasm of columnar cells. Transitional mucosa (TM) showed a decrease in DBA binding to goblet-cell mucin, which was replaced by an increase in PNA reactivity. The DBA linear pattern in the apical cytoplasm of columnar cells was unmodified, however. Changes similar to those of TM were observed in juvenile and Peutz-Jeghers polyps. Adenomas showed a progressive loss of DBA reactivity and an increase in PNA positivity related to the degree of dysplasia. This change was more evident in the linear pattern of apical cytoplasm. Only 32% of the carcinomas reacted with DBA and those were mucinous and well-differentiated adenocarcinomas. WGA was positive in all carcinomas with a different pattern than in normal mucosa. These findings suggest that the different lectin-binding patterns in normal and neoplastic colonic mucosa are related to the degree of cellular differentiation. In the process of malignant transformation the carbohydrate distribution undergoes progressive changes through the adenoma-carcinoma sequence. These changes are related to the degree of dysplasia in adenomas and to the degree of differentiation in carcinomas.

  1. Anthocyanin Content in Seeds, Leaves and Flowers of Lablab Purpureus

    USDA-ARS?s Scientific Manuscript database

    Lablab purpureus contain bioactive phytochemicals and with potential to be utilized in the pharmaceutical and nutraceutical markets. Ninety four Lablab purpureus accessions are conserved at the USDA, ARS, Plant Genetic Resources Conservation Unit in Griffin, GA. Anthocyanins are present in flowers...

  2. Influence of maize/lablab intercropping on lepidopterous stem borer infestation in maize.

    PubMed

    Maluleke, Mary H; Addo-Bediako, Abraham; Ayisi, Kingsley K

    2005-04-01

    Lepidopterous stem borers seriously affect production of maize, Zea mays L., in sub-Saharan Africa. Intercropping maize with legumes such as lablab, Lablab purpurens (L.), is one of the effective systems to control stem borers. Sole culture maize and maize/lablab intercrop system of different lablab densities were planted at two locations to investigate the effects of intercrop system on incidence and severity of stem borers with particular reference to Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae). Stem borer infestation was found to be more severe in sole culture maize than maize in maize/lablab intercrop. There was a significantly negative relationship between lablab densities and maize grain yields, suggesting a possible competition for resources between the two crops. It was concluded that density of lablab and date of planting of lablab in maize/lablab intercropping have significant affects on stem borer populations and maize grain yields.

  3. Lablab purpureus-A Crop Lost for Africa?

    PubMed

    Maass, Brigitte L; Knox, Maggie R; Venkatesha, S C; Angessa, Tefera Tolera; Ramme, Stefan; Pengelly, Bruce C

    2010-09-01

    In recent years, so-called 'lost crops' have been appraised in a number of reviews, among them Lablab purpureus in the context of African vegetable species. This crop cannot truly be considered 'lost' because worldwide more than 150 common names are applied to it. Based on a comprehensive literature review, this paper aims to put forward four theses, (i) Lablab is one of the most diverse domesticated legume species and has multiple uses. Although its largest agro-morphological diversity occurs in South Asia, its origin appears to be Africa. (ii) Crop improvement in South Asia is based on limited genetic diversity. (iii) The restricted research and development performed in Africa focuses either on improving forage or soil properties mostly through one popular cultivar, Rongai, while the available diversity of lablab in Africa might be under threat of genetic erosion. (iv) Lablab is better adapted to drought than common beans (Phaseolus vulgaris) or cowpea (Vigna unguiculata), both of which have been preferred to lablab in African agricultural production systems. Lablab might offer comparable opportunities for African agriculture in the view of global change. Its wide potential for adaptation throughout eastern and southern Africa is shown with a GIS (geographic information systems) approach.

  4. Lablab purpureus—A Crop Lost for Africa?

    PubMed Central

    Knox, Maggie R.; Venkatesha, S. C.; Angessa, Tefera Tolera; Ramme, Stefan; Pengelly, Bruce C.

    2010-01-01

    In recent years, so-called ‘lost crops’ have been appraised in a number of reviews, among them Lablab purpureus in the context of African vegetable species. This crop cannot truly be considered ‘lost’ because worldwide more than 150 common names are applied to it. Based on a comprehensive literature review, this paper aims to put forward four theses, (i) Lablab is one of the most diverse domesticated legume species and has multiple uses. Although its largest agro-morphological diversity occurs in South Asia, its origin appears to be Africa. (ii) Crop improvement in South Asia is based on limited genetic diversity. (iii) The restricted research and development performed in Africa focuses either on improving forage or soil properties mostly through one popular cultivar, Rongai, while the available diversity of lablab in Africa might be under threat of genetic erosion. (iv) Lablab is better adapted to drought than common beans (Phaseolus vulgaris) or cowpea (Vigna unguiculata), both of which have been preferred to lablab in African agricultural production systems. Lablab might offer comparable opportunities for African agriculture in the view of global change. Its wide potential for adaptation throughout eastern and southern Africa is shown with a GIS (geographic information systems) approach. PMID:20835399

  5. Fermentability of Corn-Lablab Bean Mixtures from Different Planting Densities

    USDA-ARS?s Scientific Manuscript database

    This study was conducted to determine silage fiber characteristics and fermentation profiles of corn (Zea mays L.) grown in mixture with lablab bean [Lablab purpureus (L.) Sweet] at different planting densities. The experiment was conducted in two environments in 2005. 'Rongai' lablab bean and corn ...

  6. Arcelins from an Indian wild pulse, Lablab purpureus, and insecticidal activity in storage pests.

    PubMed

    Janarthanan, Sundaram; Suresh, Palaniappan; Radke, Gary; Morgan, Thomas D; Oppert, Brenda

    2008-03-12

    A partially purified protein fraction was isolated from seed flour of the Indian wild bean, Lablab purpureus, by ion exchange and size-exclusion chromatographies. Partially purified L. purpureus proteins had hemagglutination and glycoslyation properties similar to those of lectins or lectin-like proteins from other pulses. Data obtained from two-dimensional gel electrophoresis, MALDI-TOF, and MALDI-TOF/TOF and N-terminal protein sequencing of the isolated polypeptides from L. purpureus demonstrated that the extract contained proteins similar to isoforms of arcelins 3 and 4 and pathogenesis-related protein 1 (PvPR1) of Phaseolus vulgaris. L. purpureus proteins were resistant to degradation by the commercial enzymes trypsin and chymotrypsin and were moderately resistant to pepsin, but were readily hydrolyzed to smaller peptides by papain. Insect feeding bioassays of the extract with the storage pests Rhyzopertha dominica and Oryzaephilus surinamensis, internal and external feeders of grain, respectively, demonstrated that L. purpureus proteins at 2% in the diet resulted in retarded development. However, a 5% dose of the L. purpureus fraction resulted in complete mortality of all larvae in both species. This study has demonstrated that proteins in the partially purified L. purpureus extract have the potential to control storage pests in cereals transformed with L. purpureus defense-related genes, but the need for more studies regarding efficacy and safety is discussed.

  7. Wisconsin - Increased corn silage protein with intercropped lablab bean

    USDA-ARS?s Scientific Manuscript database

    Protein supplements for livestock are costly. In recent research in southern WI, lablab bean grown with corn increased forage CP concentration over monoculture corn without compromising forage yield or potential milk production per acre. Corn was intercropped with each of three climbing beans: lab...

  8. Lectin-binding by sporozoites of Elmeria tenella.

    PubMed

    Fuller, A L; McDougald, L R

    2002-02-01

    Sporozoites of Eimeria tenella were reacted in vitro with 19 different lectins characterized with a variety of carbohydrate-binding properties. Nine lectins caused sporozoite agglutination, which was inhibited by the specific carbohydrates mannose, sialic acid, melibiose, D-galactose, or D-galNAc. When intact live or fixed whole sporozoites were reacted with fluorescein isothiocyanate-conjugated lectins, another nine lectins bound to sporozoites, giving weak to strong fluorescence but not agglutination. Of these, all nine lectins bound to surface sites, but four also bound to the refractile body. Two of the agglutinating lectins also bound to intracellular organelles of air-dried sporozoites. SDS-PAGE analysis showed that biotinylated lectins bound a wide variety of parasite proteins. Lectins with similar carbohydrate specificities had some similarity in binding patterns of parasite proteins, as well as marked differences. In a few cases lectins with different carbohydrate specificities bound common protein bands. Only one lectin (Dolichos biflorus) showed no evidence of binding to whole sporozoites, organelles, or proteins.

  9. Effect of plant lectins on Ustilago maydis in vitro.

    PubMed

    Santiago, A P; Saavedra, E; Pérez Campos, E; Córdoba, F

    2000-12-01

    Ustilago maydis is an edible parasitic basidiomycete, which specifically infects corn (Zea mays) and teocintle (Z. diploperennis). To characterise the interaction between the basidiomycete and its host organism, we tested the effect of plant lectins with well-known sugar specificity on the growth and germination of U. maydis spores. Lectins specific for N-acetyl-D-galactosamine, such as those from Dolichos biflorus and Phaseolus lunatus, and the wheatgerm agglutinin specific for N-acetyl-D-glucosamine inhibited spore germination, but were ineffective in modifying U. maydis cell growth. The galactose-specific lectin from the corn coleoptyle inhibited both germination and cell growth, while the lectin concanavalin A (mannose/glucose specific) activated spore germination and growth. Our results suggest that specific saccharide-containing receptors participate in regulating the growth and maturation of U. maydis spores.

  10. Histochemical characterization of the lectin-binding sites in the equine vomeronasal organ.

    PubMed

    Lee, Jee-young; Kang, Tae-young; Lee, Yong-duk; Shin, Tae-kyun

    2003-04-01

    The binding specificities of various lectins, such as the Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), and the Bandeiraea simplicifolia BS-1 (Isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I) lectins, were studied in the vomeronasal organ of the horse. The microvilli of the vomeronasal sensory epithelium were positive for DBA, SBA, Isolectin B4, WGA, PNA, and UEA-I. The receptor cells showed intense reactivity for DBA and WGA. Lectins were not detected in the supporting cells or basal cells. The Jacobson's glands were positive for WGA and UEA-I, but lectins were absent from the nerve bundles. From these results, we postulate that several lectin-binding carbohydrates on the microvilli and neurosensory cells are associated with chemoreception in the horse. In addition, the differential lectin-binding patterns in the horse suggest that the carbohydrates present in this particular sense organ are species-specific.

  11. Green manure and forage potential of lablab in the U.S. Southern Plains

    USDA-ARS?s Scientific Manuscript database

    Current summer forages available to support grazing by yearling cattle in the U.S. southern Great Plains have a mid-July through September quality gap. This study tested the function of the tropical/subtropical legume lablab [Lablab purpureus (L.) Sweet] as green manure or forage in central Oklahom...

  12. Green manure and forage potential of lablab in the U.S. southern plains

    USDA-ARS?s Scientific Manuscript database

    Current perennial summer forages available to support grazing by yearling cattle in the U.S. southern Great Plains have mid-July through September quality gaps. This study tested the function of the tropical/subtropical legume lablab [Lablab purpureus (L.)Sweet]as green manure or forage in central O...

  13. Glycan profiling of endometrial cancers using lectin microarray.

    PubMed

    Nishijima, Yoshihiro; Toyoda, Masashi; Yamazaki-Inoue, Mayu; Sugiyama, Taro; Miyazawa, Masaki; Muramatsu, Toshinari; Nakamura, Kyoko; Narimatsu, Hisashi; Umezawa, Akihiro; Mikami, Mikio

    2012-10-01

    Cell surface glycans change during the process of malignant transformation. To characterize and distinguish endometrial cancer and endometrium, we performed glycan profiling using an emerging modern technology, lectin microarray analysis. The three cell lines, two from endometrial cancers [well-differentiated type (G1) and poorly differentiated type (G3)] and one from normal endometrium, were successfully categorized into three independent groups by 45 lectins. Furthermore, in cancer cells, a clear difference between G1 and G3 type was observed for the glycans recognized with six lectins, Ulex europaeus agglutinin I (UEA-I), Sambucus sieboldiana agglutinin (SSA), Sambucus nigra agglutinin (SNA), Trichosanthes japonica agglutinin I (TJA-I), Amaranthus caudatus agglutinin (ACA), and Bauhinia purpurea lectin (BPL). The lectin microarray analysis using G3 type tissues demonstrated that stage I and stage III or IV were distinguished depending on signal pattern of three lectins, Dolichos biflorus agglutinin (DBA), BPL, and ACA. In addition, the analysis of the glycans on the ovarian cancer cells showed that only anticancer drug-sensitive cell lines had almost no activities to specific three lectins. Glycan profiling by the lectin microarray may be used to assess the characteristics of tumors and potentially to predict the success of chemotherapy treatment.

  14. Lectins influence chondrogenesis and osteogenesis in limb bud mesenchymal cells.

    PubMed

    Talaei-Khozani, Tahereh; Monsefi, Malihezaman; Ghasemi, Mansoureh

    2011-02-01

    The role of cell surface glycoproteins in cell behavior can be characterized by their interactions with plant lectins. This study was designed to identify the effects of lectins on chondrogenesis and osteogenesis in limb bud mesenchymal cells in vitro. Limb bud mesenchymal cells from mouse embryos were cultured in high-density micromass culture. Wheat germ agglutinin (WGA), concanavalin A (ConA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA) and Ricinus communis agglutinin (RCA) were added separately to the culture media. Cells were cultured for 5 or 9 days, and cell viability was assayed by neutral red on day 5. The micromasses were stained with alcian blue, alizarin red S and Von Kossa stains, and alkaline phosphatase assays were also done. Dolichos biflorus agglutinin induced an increase in chondrogenesis, calcium precipitation and proteoglycan production. ConA and PNA did not affect chondrocyte differentiation but induced chondrocytes to produce more proteoglycan. Wheat germ agglutinin reduced chondrification and ossification but induced mesenchymal cells to store lipid droplets. Ricinus communis agglutinin 1 was toxic and significantly reduced cell survival. In conclusion, DBA was the most effective inducer of ossification and chondrification. Wheat germ agglutinin induced adipogenesis instead. These assays showed that lectins play important roles in limb bud development.

  15. Species differences in lectin binding to pulmonary cells: Soybean agglutinin (SBA) as a marker of type I alveolar epithelial cells and alveolar macrophages in mini pigs.

    PubMed

    Kasper, M; Haroske, G; Müller, M

    1994-03-01

    We compared lectin staining patterns in rat and mini pig tissues of normal and fibrotic (irradiation-induced) lungs. Two lectins were studied: Dolichos biflorus (DBA) and Soybean (SBA). Both lectins strongly stained a subpopulation of alveolar macrophages. In the rat, DBA positive macrophages were a subpopulation of the SBA binding cells. In mini pig lungs, a further specific binding of DBA and SBA was observed: DBA reacted with endothelia, and SBA stained the alveolar type I cells. Double immunofluorescence experiments using a type II cell-specific cytokeratin antibody confirmed the selective reactivity of SBA with type I cells, which was also present in fibrotic areas with epithelial cell proliferation.

  16. Lectin binding patterns in the vomeronasal organ and accessory olfactory bulb of the rat.

    PubMed

    Salazar, I; Sánchez Quinteiro, P

    1998-10-01

    A number of previous studies have indicated that lectin histochemistry is an obvious choice for characterizing the vomeronasal system. However, apparently inconsistent results have been obtained: notably, the affinity with which various lectins bind to the accessory olfactory bulb varies among taxa, even considering closely related species. In the present study, the binding patterns of seven lectins in the rat accessory olfactory bulb, vomeronasal nerves and vomeronasal duct were investigated. The Bandeiraea simplicifolia lectin bound exclusively to the vomeronasal nerve and glomerular layers of the accessory olfactory bulb, while the Ulex europeus and Lycopersicon esculentum lectins bound to these regions and additionally to the nerve and glomerular layers of the main olfactory bulb. Soybean agglutinin showed a similar pattern to that obtained with the Ulex europeus and Lycopersicon esculentum lectins, though it also faintly labelled other parts of the structures examined. The Vicia villosa and Erythrina cristagalli lectins were not specific for the vomeronasal system, since they labelled grey and white matters in structures including the lateral olfactory tract and the anterior olfactory nuclei. The Dolichos biflorus lectin did not bind to vomeronasal tissues. The observed patterns of binding in the accessory olfactory bulb were consistent with those observed in the vomeronasal nerves, but unlike those observed in the epithelium of the vomeronasal duct. This latter result probably reflects binding of lectins to sugar residues contained in secreted mucus rather than those in epithelial nerve endings.

  17. Effects of Feeding Corn-lablab Bean Mixture Silages on Nutrient Apparent Digestibility and Performance of Dairy Cows.

    PubMed

    Qu, Yongli; Jiang, Wei; Yin, Guoan; Wei, Chunbo; Bao, Jun

    2013-04-01

    This study estimated the fermentation characteristics and nutrient value of corn-lablab bean mixture silages relative to corn silages. The effects of feeding corn-lablab bean mixture silages on nutrient apparent digestibility and milk production of dairy cows in northern China were also investigated. Three ruminally cannulated Holstein cows were used to determine the ruminal digestion kinetics and ruminal nutrient degradability of corn silage and corn-lablab bean mixture silages. Sixty lactating Holstein cows were randomly divided into two groups of 30 cows each. Two diets were formulated with a 59:41 forage: concentrate ratio. Corn silage and corn-lablab bean mixture silages constituted 39.3% of the forage in each diet, with Chinese wildrye hay constituting the remaining 60.7%. Corn-lablab bean mixture silages had higher lactic acid, acetic acid, dry matter (DM), crude protein (CP), ash, Ca, ether extract concentrations and ruminal nutrient degradability than monoculture corn silage (p<0.05). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) concentrations of corn-lablab bean mixture silages were lower than those of corn silage (p<0.05). The digestibility of DM, CP, NDF, and ADF for cows fed corn-lablab bean mixture silages was higher than for those fed corn silage (p<0.05). Feeding corn-lablab bean mixture silages increased milk yield and milk protein of dairy cows when compared with feeding corn silage (p<0.05). The economic benefit for cow fed corn-lablab bean mixture silages was 8.43 yuan/day/cow higher than that for that fed corn silage. In conclusion, corn-lablab bean mixture improved the fermentation characteristics and nutrient value of silage compared with monoculture corn. In this study, feeding corn-lablab bean mixture silages increased milk yield, milk protein and nutrient apparent digestibility of dairy cows compared with corn silage in northern China.

  18. Effects of Feeding Corn-lablab Bean Mixture Silages on Nutrient Apparent Digestibility and Performance of Dairy Cows

    PubMed Central

    Qu, Yongli; Jiang, Wei; Yin, Guoan; Wei, Chunbo; Bao, Jun

    2013-01-01

    This study estimated the fermentation characteristics and nutrient value of corn-lablab bean mixture silages relative to corn silages. The effects of feeding corn-lablab bean mixture silages on nutrient apparent digestibility and milk production of dairy cows in northern China were also investigated. Three ruminally cannulated Holstein cows were used to determine the ruminal digestion kinetics and ruminal nutrient degradability of corn silage and corn-lablab bean mixture silages. Sixty lactating Holstein cows were randomly divided into two groups of 30 cows each. Two diets were formulated with a 59:41 forage: concentrate ratio. Corn silage and corn-lablab bean mixture silages constituted 39.3% of the forage in each diet, with Chinese wildrye hay constituting the remaining 60.7%. Corn-lablab bean mixture silages had higher lactic acid, acetic acid, dry matter (DM), crude protein (CP), ash, Ca, ether extract concentrations and ruminal nutrient degradability than monoculture corn silage (p<0.05). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) concentrations of corn-lablab bean mixture silages were lower than those of corn silage (p<0.05). The digestibility of DM, CP, NDF, and ADF for cows fed corn-lablab bean mixture silages was higher than for those fed corn silage (p<0.05). Feeding corn-lablab bean mixture silages increased milk yield and milk protein of dairy cows when compared with feeding corn silage (p<0.05). The economic benefit for cow fed corn-lablab bean mixture silages was 8.43 yuan/day/cow higher than that for that fed corn silage. In conclusion, corn-lablab bean mixture improved the fermentation characteristics and nutrient value of silage compared with monoculture corn. In this study, feeding corn-lablab bean mixture silages increased milk yield, milk protein and nutrient apparent digestibility of dairy cows compared with corn silage in northern China. PMID:25049816

  19. Lectin histochemistry of palatine glands in the developing rat.

    PubMed

    Hakami, Zaki; Kitaura, Hideki; Honma, Shiho; Wakisaka, Satoshi; Takano-Yamamoto, Teruko

    2014-05-01

    This study examined the binding pattern of lectins, soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin-I (UEA-I), peanut agglutinin (PNA), wheat germ agglutinin (WGA), and succinylated WGA (sucWGA) in the developing rat palatine glands. In adult rats, heterogeneous lectin binding patterns were revealed between the anterior and posterior portions of palatine glands, as DBA, VVA, and WGA were bound more intensely and broadly in the posterior portion. SBA, PNA, and sucWGA showed far less reactivity in the anterior than in the posterior portion. At embryonic day 18 (E18), weak labeling was observed with UEA-I and WGA at the basal membrane of terminal buds, UEA-I and PNA labeled the epithelial cord, and there was no apparent binding for SBA, DBA, VVA, and sucWGA. At E20, after acinar lumenization, all lectins were detected at the acinar cell basal membranes. After birth, all lectins detectably labeled at the mucous cell apical membranes and progressively, with maturation, extended from the apical to basal portions of the cytoplasm. Apparent serous cells were observed around postnatal day 10 (PN10) and bound UEA-I. Lectins reached peak reactivity at PN21 and the binding patterns became identical to those of adults around PN28.

  20. Architecture of Deinococcus geothermalis biofilms on glass and steel: a lectin study.

    PubMed

    Peltola, Minna; Neu, Thomas R; Raulio, Mari; Kolari, Marko; Salkinoja-Salonen, Mirja S

    2008-07-01

    Deinococcus geothermalis is resistant to chemical and physical stressors and forms tenuous biofilms in paper industry. The architecture of its biofilms growing on glass and on stainless acid proof steel was studied with confocal laser scanning microscopy and fluorescent lectins and nanobeads as in situ probes. Hydrophobic nanobeads adhered to the biofilms but did not penetrate to biofilm interior. In contrast, the biofilms were readily permeable towards many different lectins. A skeletal network of glycoconjugates, reactive with Dolichos biflorus and Maclura pomifera lectins, was prominent in the space inside the biofilm colony core but absent on the exterior. Cells in the core space of the biofilm were interconnected by a network of adhesion structures, reactive with Amaranthus caudatus lectin but with none of the 65 other tested lectins. The glycoconjugates connecting the individual cells to steel reacted with Phaseolus vulgaris lectin whereas those connecting to glass mainly reacted with A. caudatus lectin. Envelopes of all cells in the D. geothermalis biofilm reacted with several other lectins, with many different specificities. We conclude that numerous different glycoconjugates are involved in the adhesion and biofilm formation of D. geothermalis, possibly contributing its unique survival capacity when exposed to dehydration, biocidal chemicals and other extreme conditions.

  1. Interaction of lectins with membrane receptors on erythrocyte surfaces.

    PubMed

    Sung, L A; Kabat, E A; Chien, S

    1985-08-01

    The interactions of human genotype AO erythrocytes (red blood cells) (RBCs) with N-acetylgalactosamine-reactive lectins isolated from Helix pomatia (HPA) and from Dolichos biflorus (DBA) were studied. Binding curves obtained with the use of tritium-labeled lectins showed that the maximal numbers of lectin molecules capable of binding to human genotype AO RBCs were 3.8 X 10(5) and 2.7 X 10(5) molecules/RBC for HPA and DBA, respectively. The binding of one type of lectin may influence the binding of another type. HPA was found to inhibit the binding of DBA, but not vice versa. The binding of HPA was weakly inhibited by a beta-D-galactose-reactive lectin isolated from Ricinus communis (designated RCA1). Limulus polyphemus lectin (LPA), with specificity for N-acetylneuraminic acid, did not influence the binding of HPA but enhanced the binding of DBA. About 80% of LPA receptors (N-acetylneuraminic acid) were removed from RBC surfaces by neuraminidase treatment. Neuraminidase treatment of RBCs resulted in increases of binding of both HPA and DBA, but through different mechanisms. An equal number (7.6 X 10(5) of new HPA sites were generated on genotypes AO and OO RBCs by neuraminidase treatment, and these new sites accounted for the enhancement (AO cells) and appearance (OO cells) of hemagglutinability by HPA. Neuraminidase treatment did not generate new DBA sites, but increased the DBA affinity for the existing receptors; as a result, genotype AO cells increased their hemagglutinability by DBA, while OO cells remained unagglutinable. The use of RBCs of different genotypes in binding assays with 3H-labeled lectins of known specificities provides an experimental system for studying cell-cell recognition and association.

  2. Phytochemical and pharmacological studies on methanolic seeds' extract of Dolichos biflorus.

    PubMed

    Ahmad, Mansoor; Sharif, Sadaf; Mehjabeen; Sharif, Hina; Jahan, Noor; Naqvi, Ghazala Raza

    2014-03-01

    The Dolichos biflorus is a well known medicinal plant in folklore for its medicinal properties. In herbal medicine the seeds of it are mainly used as tonic, astringent, diuretic, and are also recommended in asthma, bronchitis, urinary discharges, hiccoughs, ozoena, heart trouble and other diseases of brain. The main purpose of this study is to explore and to provide experimental data on the traditional use of plant Dolichos biflorus. For this purpose we investigated the plant seed extract phytochemically and pharmacologically. Phytochemical analysis was performed on extract and powder form of the drug. Procedure use for evaluation were Identification of chemical constituent by color reaction, Fluorescence analysis of powder drug, pH (in powder and extract forms), loss on drying, Thin layer chromatography, Infrared spectroscopy, acid and saponification values. In pharmacological studies (diuretic, analgesic and anti-inflammatory activities) were tested on the extract of plant seed. The tests were carried out over albino mice taking different concentration of seed extract. Seeds extract of Dolichos biflorus has exhibited mild analgesic activity, the results were (84.6±6.68) at dose 300mg/kg and (92.2±6.81) at dose 500mg/kg which were not much significant as compared to reference drug Aspirin (300mg/kg) having result (36.4±2.27). While seed extract of Dolichos biflorus exhibited remarkable diuretic activity, the values at 300 mg/kg was (1.33±0.13) and at 500 mg/kg were (2.66±0.31) which are highly significant as compared to drug Lasix (20mg /kg) having result (2.38±0.23). Anti-inflammatory effects of crude extract of Dolichos biflorus obtained at 0.06mg/kg and 01mg/kg were (26.6±2.96) and (36±1.67) respectively. While the value for aspirin as standard drug (300mg/kg) were (17.44±1.59).This study provides a platform for further investigation for the isolation of active principles responsible for biological activity.

  3. Characterization by Lectin Histochemistry of Two Subpopulations of Parietal Cells in the Rat Gastric Glands.

    PubMed

    Gómez-Santos, Laura; Alonso, Edurne; Díaz-Flores, Lucio; Madrid, Juan F; Sáez, Francisco J

    2017-05-01

    Parietal cells undergo a differentiation process while they move from the isthmus toward the pits and the base region of the gastric gland. The aim of this work was to analyze the rat gastric glands by lectin histochemistry to show the glycans expressed by upper (young) and lower (old) parietal cells. We used lectins recognizing the most frequent sugar moieties in mammals. Each lectin was assayed alone and in combination with several deglycosylation pretreatments: (1) β-elimination, which removes O-linked oligosaccharides; (2) incubation with Peptide-N-glycosidase F, to remove N-linked glycans; (3) acid hydrolysis, which removes terminal sialic acid moieties; (4) methylation-saponification, to remove sulfate groups from sugar residues; and (5) glucose oxidase, a technique carried out with the lectin concanavalin A to convert glucose into gluconic acid. The lectins from Helix pomatia, Dolichos biflorus (DBA), Glycine max (soybean), Maclura pomifera, Arachis hypogaea (peanut), Bandeiraea simplicifolia (lectin I-B4), and Datura stramonium showed a different glycan expression in the parietal cells throughout the gastric gland. This difference supports that parietal cells undergo a maturation/degeneration process while the cells descend along the gland. The role of DBA as a marker of parietal cells previously reported should be taken with caution because these cells showed different reactivity for the lectin, ranging from negative to strong labeling.

  4. Phenotype and seed production among hyacinth bean (Lablab purpureus L. Sweet) accessions rescued using hydroponic techniques

    USDA-ARS?s Scientific Manuscript database

    Hyacinth bean, Lablab purpureus L. (Sweet) is a legume used as a vegetable, forage, and in home gardens as an ornamental plant. Many accessions do not flower during their juvenile period in Byron, GA. Other hyacinth bean accessions produce few seed when regenerated in the field. This study was condu...

  5. Mineral, flavonoid, and fatty acid concentrations in ten diverse Lablab purpureus (L.) sweet accessions.

    USDA-ARS?s Scientific Manuscript database

    Seeds of Hyacinth bean (Lablab purpureus [L.]) Sweet containing high concentrations of minerals, flavonoids and fatty acids may provide government agencies with a nutrient-dense and health-beneficial food for use in hunger stricken and nutrient deprived people. Seeds from ten hyacinth bean accession...

  6. Performance of hyacinth bean (Lablab purpureus (L.) in the southern Great Plains

    USDA-ARS?s Scientific Manuscript database

    Lablab is widely cultivated in parts of Africa, south and Central America, the Indian sub-continent and other regions of Asia. Though used as a grain crop, its potential as forage or green manure has been recognized in Brazil, Africa and Australia. While some cultivar development has occurred in sou...

  7. Hydroponic rescue and regeneration of Aeschynomene, Corchorus species, and Lablab purpureus (L.) sweet genetic resources

    USDA-ARS?s Scientific Manuscript database

    Aeschynomene, Corchorus species, and Lablab purpureus (L. Sweet) have uses ranging from forage, vegetables, nutraceutical, and medicinal. Many of these will not flower nor produce seed when grown under normal field conditions in Griffin or Byron, GA because of juvenility, photoperiod and freeze-sens...

  8. Lectin histochemical studies on the vomeronasal organ of the sheep.

    PubMed

    Ibrahim, Dalia; Nakamuta, Nobuaki; Taniguchi, Kazumi; Taniguchi, Kazuyuki

    2013-01-01

    The vomeronasal organ of sheep was examined using lectin histochemistry in order to compare the types and amounts of the glycoconjugates among various components of the vomeronasal sensory and non-sensory epithelia. In the vomeronasal sensory epithelium, Dolichos biflorus agglutinin (DBA) stained particular cells, located at the same level as the vomeronasal receptor cells, while the distribution, shape and number of the stained cells did not correspond to those of the vomeronasal receptor cells. Datura stramonium lectin (DSL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L) labeled the basal cells of both vomeronasal sensory and non-sensory epithelia. While, Wheat germ agglutinin (WGA), Succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL) and Ricinus communis agglutinin-I (RCA-120) labeled the basal cells of the sensory epithelium, and Bandeiraea simplicifolia lectin-I (BSL-I) stained the basal cells of the non-sensory epithelium, respectively. Seventeen lectins labeled the free border of both vomeronasal sensory and non-sensory epithelia, while Sophora japonica agglutinin (SJA), Jacalin and Pisum sativum agglutinin (PSA) labeled neither free border of the sensory nor that of non-sensory epithelia. The expression pattern of glycoconjugate was similar, but not identical, in the free border between the sensory and non-sensory epithelia. These results indicate that there are dissimilar features in the type and amount of glycoconjugates between the vomeronasal sensory and non-sensory epithelia, and at the same time, among the various cell types either in the vomeronasal sensory or non-sensory epithelium.

  9. Growth and subsequent egg production performance of shika-brown pullets fed graded levels of cooked Lablab purpureus beans.

    PubMed

    Abeke, F O; Ogundipe, S O; Sekoni, A A; Dafwvang, I I; Adeyinka, I A; Oni, O O; Abeke, A

    2007-04-01

    The lablab seeds were cooked for 30 min, sundried for three days and thereafter milled and incorporated into chickens' diets. Six treatments, comprising of six dietary levels of lablab at 0.0, 7.5, 15.0, 22.5, 30.0 and 37.5%, respectively was fed during the grower's phase. Results obtained for the growers phase indicate that final weight (g/bird), weight gain (g/bird) and feed cost (N kg(-1)) decreased significantly (p < 0.05) as the level of lablab increased in the diet. Feed intake (g/bird and g/bird/day) as well as feed cost (N/bird) and total cost (N/bird) also decreased significantly (p < 0.05) as the level oflablab increased in the diet up to 22.5% inclusion level before increasing as the level of lablab in the diets continued to increase. The level of lablab in the diet had no effect on mortality throughout the experimental period. In addition haematological parameters such as Packed Cell Volume (PCV), Haemoglobin (Hb) and Total Protein (TP) were not significantly (p < 0.05) affected by feeding diets containing graded levels of cooked lablab beans to pullets. Results obtained during the laying phase indicates that feeding lablab seed meal up to 22.5% in the diets at the growers stage had no significant adverse effect (p < 0.05) on final weight, feed intake, feed efficiency, percent henday and henhoused egg production, percent production at peak, kilogramme feed per twelve eggs, feed cost per twelve eggs, Haugh Unit and yolk index. However, age of birds at first egg, age at 50% production and age at peak egg production were increased significantly (p > 0.05) with increase in the level of lablab seed meal in the growers diets.

  10. Development of gastrointestinal surface. VIII. Lectin identification of carbohydrate differences

    SciTech Connect

    Pang, K.Y.; Bresson, J.L.; Walker, W.A.

    1987-05-01

    Binding of microvillus membranes (MVM) from newborn and adult rats by concanavalin A (Con A), Ulex europaeus (UEA I), Dolichos bifluorus (DBA), and Triticum vulgaris (WGA) was examined to determine the availability of carbohydrate-containing sites for these lectins on the intestinal surface during development. Consistent patterns of differences in the reaction of MVM with these lectins were found. Con A and UEA had much higher reactivities to MVM of adult than newborn rats. /sup 125/I-labeled-UEA gel overlay experiments revealed the abundance of UEA-binding sites in MVM of adult rat in contrast to the two binding sites in MVM of a newborn rat. DBA bound only to MVM of the adults, and very few binding sites were found in immature MVM. In contrast to these lectins, WGA binding was much higher in MVM of the newborns and decreased with maturation. Additional experiments on the age dependence of UEA and DBA reactivities revealed that the most striking changes occur in animals from 2 to 2 wk of age. In MVM from 2-wk-old rats, there were only 13.9% and < 0.2% of the adult binding capacities for UEA and DBA, respectively. By the time the animals were 4 wk old, the binding capacity for UEA had attained close to the level of the adults, whereas for DBA it reached 71.3% of the adult value. These results provide definite evidence of changes in the intestinal surface during perinatal development.

  11. Seasonal lectin binding variations of thumb pad in the frog (Pelophylax ridibundus).

    PubMed

    Kaptan, Engin; Bolkent, Sehnaz

    2014-01-01

    The thumb pad is one of the most common secondary sexual characteristics in frogs. Although it is known that amphibian skin has affinity for several lectins, there is no report regarding lectin-binding affinity of the thumb pad or its structural components. This study investigated localization and seasonal variation of specific carbohydrate moieties of glycoconjugates in both the epidermal and dermal components of the frog thumb pad at the light microscopic level using lectin histochemistry. The study consisted of four seasonal groups of the frog species, Pelophylax ridibundus (Synonym of Rana ridibunda): active, prehibernating, hibernating and posthibernating. Four horseradish peroxidase conjugated lectins were employed. It was found that dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and ulex europaeus (UEAI) gave positive reactions in both epidermal layers and breeding glands. These three lectins bound specific secretory cells in the breeding glands, and the distribution of the cells and epithelial lectin reactions exhibited seasonal changes. In addition, UEA-I and peanut agglutinin (PNA) showed an affinity in granular glands and the granular zone of mixed glands. Generally, epidermal lectin binding showed dense affinity during the posthibernation period. DBA, UEA-I, and WGA-specific cells in the mucous gland decreased gradually until the posthibernation period. These findings suggest that differences of lectin binding in the thumb pad may be related to functional activities and, thus, seasonal adaptations. Moreover, the presence of specific lectin-binding cells in the breeding glands indicated that they consisted of heterogeneous secretory cell composition or that the cells were at different secretory stages.

  12. Lectin characterization of gonococci from an outbreak caused by penicillin-resistant Neisseria gonorrhoeae.

    PubMed Central

    Schalla, W O; Rice, R J; Biddle, J W; Jeanlouis, Y; Larsen, S A; Whittington, W L

    1985-01-01

    A total of 40 Neisseria gonorrhoeae isolates, representing 19 penicillin-resistant isolates (from 8 heterosexual patients and 11 homosexual patients) and 21 penicillin-susceptible isolates (from 15 heterosexual patients and 6 homosexual patients) and obtained from the same geographic area, were examined. Lectin agglutination patterns were based on the reactivity of the isolates with the following 14 lectins: concanavalin A, Lens culinaris, Trichosanthes kinlowii, Griffonia simplicifolia I, Arachis hypogeae (peanut agglutinin), Glycine max (soybean agglutinin), Dolichos bifloris, Griffonia simplicifolia II, Solanum tuberosum (potato starch agglutinin), Triticum vulgaris (wheat germ agglutinin), Limax flavus, Phaseolus vulgaris, Ulex europaeus I, and Lotus tetragonolobus. All isolates were serotyped with monoclonal antibodies specific for gonococcal outer membrane protein I and auxotyped, and the plasmid content was determined. Resistant patient isolates were selected for their decreased penicillin susceptibility, and control isolates were selected for their penicillin susceptibility. Even though the patient isolates demonstrated resistance to penicillin, no phenotypic differences in lectin-grouping patterns were demonstrated between the two study groups; i.e., two predominant lectin groups were observed. No resistance-associated plasmids were detected. All patient isolates were serogroup IB (serovars IB-1, IB-2, and IB-4), whereas 12 of 21 control isolates were serogroup IA (P less than 0.05). Isolates obtained from different anatomical sites in the same patient (cervical and rectal) agreed with regard to lectin patterns and serovars but not auxotypes. PMID:3935658

  13. The interaction between wheat germ agglutinin and other plant lectins with prostate cancer cells Du-145.

    PubMed

    Gabor, F; Klausegger, U; Wirth, M

    2001-06-19

    The bioadhesive properties of fluorescein-labeled plant lectins with different carbohydrate specificities were investigated by flow cytometry at 4 and 37 degrees C using Du-145 prostate cancer cells. At both temperatures the lectin association rate increased following the order: Dolichos biflorus agglutinin (DBA)lectin < wheat germ agglutinin (WGA), reflecting the glycosylation pattern of Du-145 cells. Both, the BSA-binding capacity of the cells referring to nonspecific binding and inhibition studies using the complementary carbohydrate, assured specificity of the lectin-cell interactions except for DBA. The WGA-association rate of Du-145 cells was dependent on temperature indicative for cellular uptake of membrane-bound WGA. Intracellular enrichment of WGA was confirmed by confocal microscopy. As resulted from experiments in presence of ouabain active transport mechanisms were involved in cellular uptake of WGA. Equilibration of the intracellular pH with monensin pointed to accumulation of intracellular located WGA within acidic compartments of Du-145 cells such as the lysosomes or the trans-Golgi complex. Consequently the interaction of WGA with Du-145 cells at 37 degrees C is a one way process due to immediate active transport of membrane-bound lectin into acidic compartments of prostate cancer cells.

  14. Subsets of epidermal Langerhans cells as defined by lectin binding profiles.

    PubMed

    Schuler, G; Romani, N; Linert, J; Shevach, E M; Stingl, G

    1983-11-01

    In this study we characterize the cell surface glycoconjugate moieties of strain 2 guinea pig epidermal Langerhans cells (LC) in single cell suspension by using a battery of 17 fluorescent lectins. All LC displayed binding sites for concanavalin A, succinylated concanavalin A, Lens culinaris agglutinin, Pisum sativum agglutinin, wheat germ agglutinin, succinylated wheat germ agglutinin, Griffonia simplicifolia agglutinin I, Ricinus communis agglutinin I, Phaseolus vulgaris E agglutinin, and Phaseolus vulgaris L agglutinin, but failed to bind Sophora japonica agglutinin (SJA), Dolichos biflorus agglutinin (DBA), and Ulex europaeus agglutinin I (UEA I). Neuraminidase pretreatment rendered LC reactive for SJA, but not for DBA and UEA I. The binding profiles of certain lectins point to the existence of LC subpopulations in that Griffonia simplicifolia I-B4 isolectin, peanut agglutinin (PNA), Helix pomatia agglutinin, and soybean agglutinin bound to only 80% (range 70-90%) of Ia-positive epidermal cells; binding sites for these lectins on primarily unreactive Ia-positive cells were unmasked when epidermal cells were treated with neuraminidase prior to lectin labeling. Ultrastructural PNA labeling studies revealed that the vast majority of Birbeck granule-containing LC displayed PNA binding sites, whereas indeterminate cells were consistently PNA-negative. Identification of carbohydrate configurations expressed on LC surfaces by lectin binding may provide a clue for the elucidation of the mechanisms of established LC functions and possibly the discovery of as yet unknown properties of this cell type.

  15. Influence of spices on protein utilisation of winged bean (Psophocarpus tetragonolobus) and horsegram (Dolichos biflorus).

    PubMed

    Pradeep, K U; Geervani, P

    1994-10-01

    The influence of a mixture of eleven spices commonly consumed in India on the utilisation of protein from boiled winged bean (Psophocarpus tetragonolobus) and horsegram (Dolichos biflorus) was studied at 10 and 20 percent level of protein intake in experimental rats. Spices used in the mixture include red chillies (Capsicum annum), black pepper (Piper nigrum), coriander (Coriandrum sativum), cumin (Cuminum cyminum), garlic (Allium sativum), ajowan (Carum copticum), turmeric (Curcuma longa), caraway seeds (Carum carui) and fennel seeds (Foeniculum vulgare). Addition of this spice mixture at 1.5% level of the diet decreased the TD of both legumes, significantly only in the case of horsegram. A significant increase was observed in the BV of both the legumes at both levels of protein tested.

  16. Cellular heterogeneity in the membrana granulosa of developing rat follicles: assessment by flow cytometry and lectin binding.

    PubMed

    Kerketze, K; Blaschuk, O W; Farookhi, R

    1996-07-01

    The hormone-mediated maturation of ovarian follicles is apparently accompanied by position-specific differentiation of cells of the membrana granulosa. We have assessed the extent of this cellular heterogeneity by flow cytometry using a variety of fluorescein isothiocyanate-labeled lectins as probes. Follicular development was stimulated in immature rats by treatment with either diethylstilbestrol (DES) or equine CG (eCG). Lectin binding to monodispersed rat granulosa cells was then analyzed by flow cytometry. Our results demonstrate that there are two distinct populations of small (4-7 microM) and large (9-12 microM) granulosa cells in follicles from DES- and eCG-treated animals. Both populations appear to be mitotically active and show specific lectin-binding characteristics. Six lectins (canavalia ensiforms, triticum vulgaris, maclura pomifera, erythrina cristagalli, jacalin, and vicia villosa) bind equally to both small and large granulosa cells from the DES- and eCG-treated rats. In contrast, no binding to either cell population was detected with six other lectins (dolichos biflorus, griffonia simplicifolia-II, lycopersicon esculentum, datura stramonium, solanum tuberosum, and ulex europaeus). Furthermore, four galactose-binding lectins (bauhinia purpurea, glysine maximus, griffonia simplicifolia-I, and arachis hypogaea) were found to identify specific subsets of granulosa cells. Three of these lectins (bauhinia purpurea, glysine maximus, and griffonia simplicifolia-I) bind to only small granulosa cells from either DES- or eCG- treated immature rats. The fourth lectin (arachis hypogaea) identifies subpopulations of both small and large granulosa cells. Application of the four galactose-specific lectins to fixed sections of frozen ovaries demonstrated binding to the perioocyte and cumulus granulosa cells. We conclude that cellular heterogeneity exists within the follicular epithelium at various stages-specific lectin-binding sites.

  17. Rotenoids from Lablab purpureus L. and their bioefficacy against human disease vectors.

    PubMed

    Kamal, Raka; Mathur, N

    2010-11-01

    Various plant parts of Lablab purpureu s L. were collected and analyzed separately for their rotenoid content. Among the plant parts, the maximum content was in the roots and minimum in the seeds. The identity of different rotenoids was confirmed by melting point, mixed melting point, UV, and infrared spectral studies and gas-liquid chromatography. Six rotenoids (deguelin, dehydrodeguelin, rotenol, rotenone, tephrosin, and sumatrol) were isolated, identified, and quantified both in vivo and in vitro. Toxicological studies of extracts showed bioefficacy against causal agents of malaria, dracunculiasis, and amoebiasis.

  18. Developmental changes affecting lectin binding in the vomeronasal organ of domestic pigs, Sus scrofa.

    PubMed

    Park, Junwoo; Lee, Wonho; Jeong, Chanwoo; Kim, Hwangryong; Taniguchi, Kazumi; Shin, Taekyun

    2012-01-01

    This study investigated the developmental changes of glycoconjugate patterns in the porcine vomeronasal organs (VNOs) and associated glands (Jacobson's glands) from prenatal (9 weeks of gestation) and postnatal (2 days after birth) to the sexually mature stage (6 months old). The VNO of pigs (Sus scrofa) was examined using the following: Dolichos biflorus agglutinin (DBA), Bandeiraea simplicifolia agglutinin isolectin B4 (BSI-B4), Triticum vulgaris agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), and soybean agglutinin (SBA). At the fetal stage, all lectins examined were detected mainly in the free border of the vomeronasal epithelium, but few (WGA and UEA-I) and or absent in the VNO cell bodies. At the postnatal and sexually mature stages, the reactivity of some lectins, including WGA, UEA-I, DBA and SBA, were shown to increase in the VNO sensory epithelium as well as the free border. The increased reactivity of lectins as development progressed was also observed in Jacobson's gland acini. These findings suggest that binding sites of lectins, including those of WGA, UEA-I, DBA, and SBA, increase during development from fetal to postnatal growth, possibly contributing to the increased ability of chemoreception in the pig.

  19. Rescue of photoperiod/freeze-sensitive and low seed producing accessions of Lablab purpureus using hydroponic cloning and aeroponics.

    USDA-ARS?s Scientific Manuscript database

    Hyacinth bean, Lablab purpureus is a legume used as a vegetable and the USDA, ARS, PGRCU conserves 137 hyacinth bean accessions from countries worldwide. Many accessions in this collection are photoperiod and freeze-sensitive due to their flower and seed production during November through March in t...

  20. Biological activity of Dolichos biflorus L. trypsin inhibitor against lepidopteran insect pests.

    PubMed

    Nath, Amarjit K; Kumari, Reena; Sharma, Shilpa; Sharma, Heena

    2015-09-01

    Protease inhibitors confer resistance in plants against insect pests by inhibiting larval gut proteases. Cultivars of Dolichos biflorus were screened for their inhibitory activity against midgut proteases of Pieris brassicae larvae. Seed extracts of developing and germinating seeds of HPK4 cultivar inhibited larval gut proteases of Spodoptera littoralis efficiently. Neonate larvae of P. brassicae fed on cabbage leaf discs coated with 0.025-2.50 mg protein (seed extract) resulted in 10-80% larval mortality and significantly reduced leaf area eaten and faecal matter as compared to control. The treated larvae had 40% less soluble proteins per mg faecal matter and there was similar decline in midgut proteases of treated larvae (@ 2.5 mg protein) compared to untreated ones after 5 days. The LC50 and LT50 value was calculated to be 1.05 mg/leaf disc and 4.8 days (2.5 mg protein), respectively for neonate larvae of P. brassicae. Significant reduction in egg hatching (75%) was observed in egg mass treated with 5.3 mg of crude inhibitor protein of mature seeds. This could be due to the inhibition of proteases involved in the hydrolysis of egg chorion proteins. The studies demonstrated the insecticidal activity of D. biflorus seed extracts.

  1. Anthocyanins and flavonols are responsible for purple color of Lablab purpureus (L.) sweet pods.

    PubMed

    Cui, Baolu; Hu, Zongli; Zhang, Yanjie; Hu, Jingtao; Yin, Wencheng; Feng, Ye; Xie, Qiaoli; Chen, Guoping

    2016-06-01

    Lablab pods, as dietary vegetable, have high nutritional values similar to most of edible legumes. Moreover, our studies confirmed that purple lablab pods contain the natural pigments of anthocyanins and flavonols. Compared to green pods, five kinds of anthocyanins (malvidin, delphinidin and petunidin derivatives) were found in purple pods by HPLC-ESI-MS/MS and the major contents were delphinidin derivatives. Besides, nine kinds of polyphenol derivatives (quercetin, myricetin, kaempferol and apigenin derivatives) were detected by UPLC-ESI-MS/MS and the major components were quercetin and myricetin derivatives. In order to discover their molecular mechanism, expression patterns of biosynthesis and regulatory gens of anthocyanins and flavonols were investigated. Experimental results showed that LpPAL, LpF3H, LpF3'H, LpDFR, LpANS and LpPAP1 expressions were significantly induced in purple pods compared to green ones. Meanwhile, transcripts of LpFLS were more abundant in purple pods than green or yellow ones, suggestind that co-pigments of anthocyanins and flavonols are accumulated in purple pods. Under continuously dark condition, no anthocyanin accumulation was detected in purple pods and transcripts of LpCHS, LpANS, LpFLS and LpPAP1 were remarkably repressed, indicating that anthocyanins and flavonols biosynthesis in purple pods was regulated in light-dependent manner. These results indicate that co-pigments of anthocyanins and flavonols contribute to purple pigmentations of pods.

  2. Molecular diversity and phylogeny of rhizobia associated with Lablab purpureus (Linn.) grown in Southern China.

    PubMed

    Chang, Yue Li; Wang, En Tao; Sui, Xin Hua; Zhang, Xiao Xia; Chen, Wen Xin

    2011-06-01

    As an introduced plant, Lablab purpureus serves as a vegetable, herbal medicine, forage and green manure in China. In order to investigate the diversity of rhizobia associated with this plant, a total of 49 rhizobial strains isolated from ten provinces of Southern China were analyzed in the present study with restriction fragment length polymorphism and/or sequence analyses of housekeeping genes (16S rRNA, IGS, atpD, glnII and recA) and symbiotic genes (nifH and nodC). The results defined the L. purpureus rhizobia as 24 IGS-types within 15 rrs-IGS clusters or genomic species belonging to Bradyrhizobium, Rhizobium, Ensifer (synonym of Sinorhizobium) and Mesorhizobium. Bradyrhizobium spp. (81.6%) were the most abundant isolates, half of which were B. elkanii. Most of these rhizobia induced nodules on L. purpureus, but symbiotic genes were only amplified from the Bradyrhizobium and Rhizobium leguminosarum strains. The nodC and nifH phylogenetic trees defined five lineages corresponding to B. yuanmingense, B. japonicum, B. elkanii, B. jicamae and R. leguminosarum. The coherence of housekeeping and symbiotic gene phylogenies demonstrated that the symbiotic genes of the Lablab rhizobia were maintained mainly through vertical transfer. However, a putative lateral transfer of symbiotic genes was found in the B. liaoningense strain. The results in the present study clearly revealed that L. purpureus was a promiscuous host that formed nodules with diverse rhizobia, mainly Bradyrhizobium species, harboring different symbiotic genes.

  3. Lectins from edible mushrooms.

    PubMed

    Singh, Senjam Sunil; Wang, Hexiang; Chan, Yau Sang; Pan, Wenliang; Dan, Xiuli; Yin, Cui Ming; Akkouh, Ouafae; Ng, Tzi Bun

    2014-12-31

    Mushrooms are famous for their nutritional and medicinal values and also for the diversity of bioactive compounds they contain including lectins. The present review is an attempt to summarize and discuss data available on molecular weights, structures, biological properties, N-terminal sequences and possible applications of lectins from edible mushrooms. It further aims to update and discuss/examine the recent advancements in the study of these lectins regarding their structures, functions, and exploitable properties. A detailed tabling of all the available data for N-terminal sequences of these lectins is also presented here.

  4. Updated version of an interim connection space LabPQR for spectral color reproduction: LabLab.

    PubMed

    Cao, Qian; Wan, Xiaoxia; Li, Junfeng; Liang, Jingxing

    2016-09-01

    In this paper, we propose a new interim connection space (ICS) called LabLab, which is an updated version of LabPQR, to overcome the drawback that the last three dimensions of LabPQR have no definite colorimetric meanings. We extended and improved the method by which the first three dimensions of LabPQR are deduced to obtain an ICS consisting of two sets of CIELAB values under different illuminants, and the reconstructed spectra from LabLab were obtained by minimizing colorimetric errors by means of the computational formula of the CIE-XYZ tristimulus values combined with least-squares best fit. The improvement obtained from the proposed method was tested to compress and reconstruct the reflectance spectra of the 1950 Natural Color System color chips and more than 50,000 ISO SOCS color patches as well as six multispectral images acquired by multispectral image acquisition systems using 1600 glossy Munsell color chips as training samples. The performance was evaluated by the mean values of color differences between the original and reconstructed spectra under the CIE 1931 standard colorimetric observer and the CIE standard illuminants D50, D55, D65, D75, F2, F7, F11, and A as well as five multichip white LED light sources. The mean and maximum values of the root mean square errors between the original and reconstructed spectra were also calculated. The experimental results show that the proposed three LabLab interim connection spaces significantly outperform principal component analysis, LabPQR, XYZLMS, Fairman-Brill, and LabRGB in colorimetric reconstruction accuracy at the cost of slight reduction of spectral reconstruction accuracy and illuminant independence of color differences of the suggested LabLab interim connection spaces outperform other interim connection spaces. In addition, the presented LabLab interim connection spaces could be quite compatible with the extensively used colorimetric management system since each dimension has definite colorimetric

  5. Postnatal morphological and lectin histochemical observation of the submucosal glands of rat nasopharynx.

    PubMed

    Hakami, Zaki; Wakisaka, Satoshi

    2016-09-01

    The development of submucosal glands of rat nasopharynx was studied with respect to their morphological maturation and glycoprotein alterations during the postnatal period. This study examined the histological morphology with hematoxylin-eosin and the binding pattern of lectins, soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin-I (UEA-I), peanut agglutinin (PNA), wheat germ agglutinin (WGA), and succinylated WGA (sucWGA) on frozen sections from newborn into adulthood. At birth, nasopharyngeal glands consisted of rudimentary secretory units which by postnatal day 3 (PN3) showed the characteristic features of salivary glands comprised of mixed mucous and serous cells. With maturation, serous cells increased in number and were arranged in clusters. Lectin reactivity at birth was detected at the acinar cell basal membranes for DBA, SBA, VVA, UEA-1 and PNA. At PN3, lectins labeled the apical cytoplasm and basolateral membranes of mucous cells and progressively with maturation, extended from the apical to basal portions of the cytoplasm with variable reactivity of VVA, PNA and sucWGA. Serous cells were labeled by UEA-1 starting from PN10 and also by PNA in adults. Ducts showed variable lectin reaction on the luminal membrane with strong reactivity of DBA and UEA-1 at PN21. Taken together, lectin histochemistry indicated the transitional occurrence of glycoproteins depending on the stage of maturation of the glands. Moreover, these results emphasize the difference in the morphology and lectin histochemistry between the nasopharyngeal and palatine glands. Copyright © 2016 Elsevier GmbH. All rights reserved.

  6. Modification of lectin binding in rat gut mucosa during experimental cholestasis.

    PubMed Central

    Vaccaro, R; Casu, C; Renda, T

    1992-01-01

    Glycoconjugate distribution on rat gut mucosa has been studied by using peroxidase-labelled lectins (Lotus tetragonolobus, Dolichos biflorus, Arachis hypogaea and Glycine max) after surgical interruption of the common bile duct. Specimens from cholestatic rats were compared with sham-operated (simple laparotomy) and normal controls to determine which of the observed modifications could be due either to the operation itself or to the cholestasis. Most of the modifications occurred in the duodenum. The operation itself modified some binding properties. Lotus tetragonolobus binding extended both in cholestatic and in sham-operated rats, but returned to normal levels earlier in sham-operated than in cholestatic rats. Conversely, cholestasis induced (1) almost total loss of Arachis hypogaea binding in the Golgi zone of superficial duodenal goblet cells; (2) an increase at the 14th postoperative day of Dolichos biflorus binding in the cytoplasmic calyx of goblet cells which then diminished up until the 28th day; and (3) an increase of Glycine max binding in the Golgi zone of goblet cells. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 PMID:1295862

  7. Cyborg lectins: novel leguminous lectins with unique specificities.

    PubMed

    Yamamoto, K; Maruyama, I N; Osawa, T

    2000-01-01

    Bauhinia purpurea lectin (BPA) is one of the beta-galactose-binding leguminous lectins. Leguminous lectins contain a long metal-binding loop, part of which determines their carbohydrate-binding specificities. Random mutations were introduced into a portion of the cDNA coding BPA that corresponds to the carbohydrate-binding loop of the lectin. An library of the mutant lectin expressed on the surface of lambda foo phages was screened by the panning method. Several phage clones with an affinity for mannose or N-acetylglucosamine were isolated. These results indicate the possibility of making artificial lectins (so-called "cyborg lectins") with distinct and desired carbohydrate-binding specificities.

  8. Histochemistry of six lectins in the tissues of the flat fish Paralichthys olivaceus.

    PubMed

    Jung, Kyung-Sook; Ahn, Mee-Jung; Lee, Yong-Duk; Go, Gyung-Min; Shin, Tae-Kyun

    2002-12-01

    Lectins are glycoproteins that specifically bind carbohydrate structures and may participate in the biodefense mechanisms of fish. In this study, the binding of three lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA) and Ulex europaeus (UEA-I) were studied in the gill, liver, intestine, kidney, heart, and spleen of the flat fish Paralichthys olivaceus. DBA was detected in intestinal mucous cells, as well as in gill epithelial and mucous cells. It was weakly detected in renal tubule epithelial cells and in bile duct epithelial cells. The strong SBA staining was seen in the intestinal club cells, in bile duct epithelial cells and renal tubule epithelial cells. There were intense positive reactions for isolectin B4 in gill epithelial and mucous cells, and the strong isolectin B4 staining was seen in epithelial cells of the bile duct and intestine. The strong WGA staining was seen in the gill mucosal cells, sinusoid, renal tubule epithelial cells and mucosal cells of the intestine. UEA-I was detected in the gill epithelial and mucosal cells, bile duct epithelial cells and renal tubular epithelial cells. These results suggest that the six lectins examined were localized in the covering epithelia of the various organs of the flat fish and they may participate in the biodefense mechanism of the intra body surface in which is exposed to various antigens.

  9. Distribution of lectin-bindings in the testis of the lesser mouse deer, Tragulus javanicus.

    PubMed

    Agungpriyono, S; Kurohmaru, M; Kimura, J; Wahid, A H; Sasaki, M; Kitamura, N; Yamada, J; Fukuta, K; Zuki, A B

    2009-06-01

    The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.

  10. Alterations in lectin binding to the epidermis following treatment with 8-methoxypsoralen plus long-wave ultraviolet radiation

    SciTech Connect

    Danno, K.; Takigawa, M.; Horio, T.

    1984-02-01

    The alterations in lectin fluorescence stainings to the epidermis were examined in guinea pig skin treated with topical application of a 1% 8-methoxypsoralen (8-MOP) solution plus long-wave ultraviolet (UVA) radiation (1.5-3.5 J/cm2) (PUVA). Serial biopsy specimens taken up to 21 days postirradiation were stained with 8 commercially available lectins labeled with either fluorescein isothiocyanate (FITC) or biotin (followed by avidin D-FITC): Bandeiraea simplicifolia agglutinin I (BSA), concanavalin A (Con-A), Dolichos biflorus agglutinin (DBA), peanut agglutinin (PNA), Ricinus communis agglutinin I (RCA), soybean agglutinin (SBA), Ulex europeus agglutinin I (UEA), and wheat germ agglutinin (WGA). In normal guinea pig skin UEA staining was absent. Following PUVA treatment, UEA and DBA stainings became apparent or stronger in intensity after days 7-14 (UEA) and days 4-7 (DBA), respectively, and returned to negative or weak by days 14-21. Stainings with Con-A, SBA, and WGA gave remarkable decreases in intensity after days 2-4 and recovered to the baseline by days 7-14. Intensity of BSA, PNA, and RCA stainings was decreased to a lesser degree than the other lectins. Such changes were not produced by application of 8-MOP, UVA radiation (less than 10 J/cm2), UVB radiation (900-2700 mJ/cm2), or tape stripping. These results suggest that PUVA treatment perturbs the composition or organization of epidermal cell surface glycoconjugates to induce alterations in lectin stainings.

  11. Lectin binding properties of liver, small intestine and tail of metamorphosing marsh frog (Pelophylax ridibundus Pallas 1771).

    PubMed

    Kaptan, Engin; Sengezer Inceli, Meliha; Sancar Bas, Serap

    2013-07-01

    In this present study, localization and variations of specific sugar moieties in the terminal carbohydrate chains of glycoconjugates in the small intestine, liver and tail have been investigated during the metamorphosis of Pelophylax ridibundus larvae. For this purpose, four lectins were used: wheat germ agglutinin (WGA), Ulex europaeus agglutinin (UEA-I), Dolichos biflorus agglutinin (DBA) and peanut agglutinin (PNA), in different larval stages of the frog. Some cells stained specifically in the intestinal mucosa and in tail epidermal cells with the lectins and their affinity changed during metamorphic transformation. For the most part, they decreased in the climax and postmetamorphic periods. It was also found that WGA, DBA and UEA-I lectins exhibited strong affinity to white blood cells in the liver and their binding affinities were the highest in prometamorphosis and they gradually decreased until the end of metamorphosis. These results suggest that the changes of lectin binding in metamorphosis may be an indication of some cellular events occurring in larval metamorphosis such as cell differentiation and damage of cell adhesion between death and differentiating cells. They also can be useful markers for detection of white blood cells in amphibian hematopoietic organs.

  12. Dolichos falcata Klein attenuated the inflammation induced by monosodium urate crystals in vivo and in vitro.

    PubMed

    Chen, Lvyi; Mola, Mahmutjian; Deng, Xukun; Mei, Zhinan; Huang, Xianju; Shu, Guangwen; Wei, Liuhong; Hou, Xiaomin; Lan, Zhou; Lin, Qinxiong

    2013-11-25

    Dolichos falcata Klein (DF), a Chinese Dai ethnic medicine popularly known as "Tuoyeteng" in Yunnan province of China, has been widely used as a traditional herbal medicine for the treatment of fracture and beriberoid disease for a long time in China. The present study was carried out to investigate the anti-inflammatory activity and the bioactive chemical constituents of DF, and further to assess its possible mechanism on gouty arthritis in an animal model of the MSU crystals-induced gouty inflammation. The ethanol extract (EE) of DF at the doses of 10, 20 and 40 mg/kg was administered to the rats treated with MSU crystals to evaluate the anti-gouty arthritis effect. Subsequently, the components of EE were isolated and identified using classical methods. Phyto-chemical analysis of EE was further carried out by HPLC-DAD. Finally, the anti-inflammatory effect of EE and two isolated components were assessed using the MSU crystals-treated monocyte/macrophage cell line RAW 264.7 in vitro. EE (10, 20 and 40 mg/kg) significantly attenuated the pain threshold value, the joint swelling degree, the inflammatory cell infiltration of articular tissue and the increased levels of pro-inflammatory cytokines in MSU crystals-treated rats. Moreover, doliroside A (DA) and medicagenic acid-3-O-β-D-glucopyranoside (MG) were isolated and identified from EE. The major components of EE, including DA, MG and other triterpenoids, were well confirmed by HPLC. A further study revealed that EE, DA and MG (10, 20, 40μg/mL) exhibited stronger inhibitory effects on the production of pro-inflammatory cytokines (including interleukin-1β, interleukin-6 and tumor necrosis factor-α) in MSU crystals-treated RAW 264.7 cells. These findings indicate that the major triterpenoids present in DF have a remarkable effect on improving symptoms of acute gouty arthritis induced by MSU crystals through inhibiting the production of pro-inflammatory cytokines. © 2013 Elsevier Ireland Ltd. All rights reserved.

  13. Lectins with potential for anti-cancer therapy.

    PubMed

    Yau, Tammy; Dan, Xiuli; Ng, Charlene Cheuk Wing; Ng, Tzi Bun

    2015-02-26

    This article reviews lectins of animal and plant origin that induce apoptosis and autophagy of cancer cells and hence possess the potential of being developed into anticancer drugs. Apoptosis-inducing lectins encompass galectins, C-type lectins, annexins, Haliotis discus discus lectin, Polygonatum odoratum lectin, mistletoe lectin, and concanavalin A, fucose-binding Dicentrarchus labrax lectin, and Strongylocentrotus purpuratus lectin, Polygonatum odoratum lectin, and mistletoe lectin, Polygonatum odoratum lectin, autophagy inducing lectins include annexins and Polygonatum odoratum lectin.

  14. Lectins of living organisms. The overview.

    PubMed

    Lakhtin, Vladimir; Lakhtin, Mikhail; Alyoshkin, Vladimir

    2011-12-01

    Occurrence, organization, and functioning of lectins as well as their current classifications are under investigation. Results indicate importance of symbiotic lectins for clinical microecology. Lectins and lectin-based approaches have wide perspectives for medical biotechnology. Lectin terms, relationships between lectins and enzymes are discussed. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Lectins: production and practical applications

    PubMed Central

    2010-01-01

    Lectins are proteins found in a diversity of organisms. They possess the ability to agglutinate erythrocytes with known carbohydrate specificity since they have at least one non-catalytic domain that binds reversibly to specific monosaccharides or oligosaccharides. This articles aims to review the production and practical applications of lectins. Lectins are isolated from their natural sources by chromatographic procedures or produced by recombinant DNA technology. The yields of animal lectins are usually low compared with the yields of plant lectins such as legume lectins. Lectins manifest a diversity of activities including antitumor, immunomodulatory, antifungal, HIV-1 reverse transcriptase inhibitory, and anti-insect activities, which may find practical applications. A small number of lectins demonstrate antibacterial and anti-nematode activities. PMID:20890754

  16. Histochemical staining using lectin probes.

    PubMed

    Akimoto, Yoshihiro; Kawakami, Hayato

    2014-01-01

    In histochemistry and cytochemistry, lectins are often used as probes for the localization of carbohydrates in cells and tissues. With lectins, cells and tissues can be identified as a particular type or a group in situ. Various lectins have been used for mapping of normal cells and tissues, pathological diagnosis such as malignant transformation, and identification of cell lineages during development. This chapter describes light and electron microscopic methods using lectin probes for determining carbohydrate localization in cells and tissues.

  17. Lectins with anti-HIV activity: a review.

    PubMed

    Akkouh, Ouafae; Ng, Tzi Bun; Singh, Senjam Sunil; Yin, Cuiming; Dan, Xiuli; Chan, Yau Sang; Pan, Wenliang; Cheung, Randy Chi Fai

    2015-01-06

    Lectins including flowering plant lectins, algal lectins, cyanobacterial lectins, actinomycete lectin, worm lectins, and the nonpeptidic lectin mimics pradimicins and benanomicins, exhibit anti-HIV activity. The anti-HIV plant lectins include Artocarpus heterophyllus (jacalin) lectin, concanavalin A, Galanthus nivalis (snowdrop) agglutinin-related lectins, Musa acuminata (banana) lectin, Myrianthus holstii lectin, Narcissus pseudonarcissus lectin, and Urtica diocia agglutinin. The anti-HIV algal lectins comprise Boodlea coacta lectin, Griffithsin, Oscillatoria agardhii agglutinin. The anti-HIV cyanobacterial lectins are cyanovirin-N, scytovirin, Microcystis viridis lectin, and microvirin. Actinohivin is an anti-HIV actinomycete lectin. The anti-HIV worm lectins include Chaetopterus variopedatus polychaete marine worm lectin, Serpula vermicularis sea worm lectin, and C-type lectin Mermaid from nematode (Laxus oneistus). The anti-HIV nonpeptidic lectin mimics comprise pradimicins and benanomicins. Their anti-HIV mechanisms are discussed.

  18. Lectin affinity electrophoresis.

    PubMed

    Kobayashi, Yuka

    2014-01-01

    An interaction or a binding event typically changes the electrophoretic properties of a molecule. Affinity electrophoresis methods detect changes in the electrophoretic pattern of molecules (mainly macromolecules) that occur as a result of biospecific interactions or complex formation. Lectin affinity electrophoresis is a very effective method for the detection and analysis of trace amounts of glycobiological substances. It is particularly useful for isolating and separating the glycoisomers of target molecules. Here, we describe a sensitive technique for the detection of glycoproteins separated by agarose gel-lectin affinity electrophoresis that uses antibody-affinity blotting. The technique is tested using α-fetoprotein with lectin (Lens culinaris agglutinin and Phaseolus vulgaris agglutinin)-agarose gels.

  19. Tracheobronchial epithelium of the sheep: IV. Lectin histochemical characterization of secretory epithelial cells.

    PubMed

    Mariassy, A T; Plopper, C G; St George, J A; Wilson, D W

    1988-09-01

    Conventional histochemical characterization of the mucus secretory apparatus is often difficult to reconcile with the biochemical analysis of respiratory secretions. This study was designed to examine the secretory glycoconjugates in airways using lectins with biochemically defined affinities for main sugar residues of mucus. We used five biotinylated lectins--DBA (Dolichos biflorus) and SBA (Glycine max) for N-acetyl galactosamine (galNAc), BSA I (Bandeiraea simplicifolia) and PNA (Arachis hypogea) for galactose (gal), and UEA I (Ulex europeus)--for detection of fucose (fuc) in HgCl2-fixed, paraffin-embedded, serially sectioned trachea, lobar and segmental bronchi and bronchioles of nine sheep. Lectins selectively localized the carbohydrate residues in luminal secretions, on epithelial cell surfaces, and in secretory cells. In proximal airways, the major carbohydrate residues in luminal secretions, cell surfaces, goblet cells, and glands were fuc and gal-NAc. PNA reacted mainly with apical granules of less than 10% of goblet cells, and gal residues were only detected in some of the mucous cells and on basolateral cell surfaces. Distal airways contained sparse secretion in the lumen, mucous cells contained weakly reactive fuc and gal-NAc, and the epithelial surfaces of Clara cells contained gal. Sugars abundant in the airway secretions were also the major component of cells in glands. We conclude that there is a correlation between specific sugar residues in secretory cells, glycocalyx, and luminal secretions in proximal and distal airways. This suggests that lectins may be used to obtain information about airway secretory cell composition from respiratory secretions.

  20. Lectin typing of Campylobacter isolates.

    PubMed Central

    O'Sullivan, N; Benjamin, J; Skirrow, M B

    1990-01-01

    Isolates of Campylobacter jejuni, C coli, C fetus and C laridis were tested for agglutination reactions with a panel of five lectins: Arachis hypogaea, Bauhinia purpurea, Solanum tuberosum, Triticum vulgaris and Wisteria floribunda. Twenty three patterns of agglutination (lectin types) were recorded among 376 isolates. Patterns were consistent and reproducible. Only 4.5% of isolates were untypable because of autoagglutination. Some lectin types were found exclusively or predominantly in a species, but others were shared between species. Forty two per cent of C jejuni and 35% of C coli isolates belonged to lectin type 4. There was no apparent correlation between lectin type and serotype; different lectin types were found among strains of single Penner and Lior serotypes. Lectin typing is a simple and economical procedure suitable for use in non-specialist laboratories, either as an adjunct to serogrouping or, after further development, as a sole typing scheme. PMID:2262570

  1. Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus

    PubMed Central

    Ko, Sang-Mu; Kwon, Joseph; Vaidya, Bipin; Choi, Jong Soon; Lee, Hee-Min; Oh, Myung-Joo; Bae, Hyeun-Jong; Cho, Se-Young; Oh, Kyung-Seo; Kim, Duwoon

    2014-01-01

    The accuracy and sensitivity of PCR-based methods for detection of hepatitis A virus (HAV) are dependent on the methods used to separate and concentrate the HAV from the infected cells. The pH and ionic strength affect the binding affinity of the virus to cells. In this study, we initially investigated the effects of pH (4.0–10.0) and metal ions (Fe2+, Co2+, Cu2+, Mg2+, K+, and Ca2+) on the binding of HAV to oyster digestive cells. The lowest relative binding (RB) of HAV to the cells was found at pH 4.0 and in FeSO4 solution (64.6% and 68.1%, respectively). To develop an alternative to antibody-dependent immunomagnetic separation prior to detection of HAV using RT-PCR, the binding of HAV to five lectins, peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Ulex europaeus agglutinin (UEA-1) and soybean agglutinin (SBA), was evaluated using ELISAs. SBA showed significantly higher RB to HAV than the other lectins tested. In addition, HAV could be concentrated within 30 min using SBA-linked magnetic bead separation (SMS) prior to the RT-PCR assay. Our findings demonstrate the feasibility of using SMS combined with RT-PCR to detect HAV at dilutions ranging from 10−1–10−4 of a HAV stock (titer: 104 TCID50/mL). PMID:24599279

  2. Comparative study of blood group-recognizing lectins toward ABO blood group antigens on neoglycoproteins, glycoproteins and complex-type oligosaccharides.

    PubMed

    Matsui, T; Hamako, J; Ozeki, Y; Titani, K

    2001-02-16

    Binding specificities of ABO blood group-recognizing lectins toward blood group antigens on neoglycoproteins, glycoproteins and complex-type oligosaccharides were studied by lectin-blotting analysis, enzyme linked immunosorbent assay and lectin-conjugated agarose column chromatography. Human serum albumin conjugated with A- and B-trisaccharides was clearly recognized by Helix pomatia (HPA), Phaseolus lunatus, Dolichos biflorus agglutinins, and Griffonia simplicifolia I agglutinin B(4), respectively. Almost the same results were obtained for human group A and B ovarian cyst and A-active hog gastric mucins, but Glycine max agglutinin only reacted to the group A hog mucin. When human plasma von Willebrand factor (vWF), having Asn-linked blood group antigens, was tested, HPA was highly sensitive to blood group A antigen on the vWF. Ulex europaeus agglutinin I (UEA-I) preferentially bound to the vWF from blood group O plasma. Within the GalNAc-recognizing lectins examined, a biantennary complex-type oligosaccharide having the blood group A structure retarded on an HPA-agarose column, and the affinity was diminished after digestion with alpha-N-acetylgalactosaminidase. This product bound to UEA-I agarose column. These results indicate that HPA and UEA-I are most sensitive for detection of glycoproteins possessing small amounts of blood group A and H antigens and also useful for fractionation of complex-type oligosaccharides with blood group A and H antigens, respectively.

  3. Agrobotanical attributes, nitrogen-fixation, enzyme activities and nutraceuticals and tyrosinase enzyme of hyacinth bean (Lablab purpureus L.) - a bio-functional medicinal legume.

    USDA-ARS?s Scientific Manuscript database

    Hyacinth bean (Lablab purpureus L.) accessions of different origins received from USDA, ARS, Plant Genetic Resources Conservation Unit, Griffin, GA, U.S.A. were evaluated for agrobotanical attributes, enzyme activities, nutraceuticals and quality in pot culture at AMU, Aligarh, Uttar Pradesh. Fresh ...

  4. Lectins from Mycelia of Basidiomycetes

    PubMed Central

    Nikitina, Valentina E.; Loshchinina, Ekaterina A.; Vetchinkina, Elena P.

    2017-01-01

    Lectins are proteins of a nonimmunoglobulin nature that are capable of specific recognition of and reversible binding to the carbohydrate moieties of complex carbohydrates, without altering the covalent structure of any of the recognized glycosyl ligands. They have a broad range of biological activities important for the functioning of the cell and the whole organism and, owing to the high specificity of reversible binding to carbohydrates, are valuable tools used widely in biology and medicine. Lectins can be produced by many living organisms, including basidiomycetes. Whereas lectins from the fruit bodies of basidiomycetes have been studied sufficiently well, mycelial lectins remain relatively unexplored. Here, we review and comparatively analyze what is currently known about lectins isolated from the vegetative mycelium of macrobasidiomycetes, including their localization, properties, and carbohydrate specificities. Particular attention is given to the physiological role of mycelial lectins in fungal growth and development. PMID:28640205

  5. Glycan and lectin biosensors

    PubMed Central

    Belický, Štefan; Katrlík, Jaroslav

    2016-01-01

    A short description about the importance of glycan biorecognition in physiological (blood cell type) and pathological processes (infections by human and avian influenza viruses) is provided in this review. Glycans are described as much better information storage media, compared to proteins or DNA, due to the extensive variability of glycan structures. Techniques able to detect an exact glycan structure are briefly discussed with the main focus on the application of lectins (glycan-recognising proteins) in the specific analysis of glycans still attached to proteins or cells/viruses. Optical, electrochemical, piezoelectric and micromechanical biosensors with immobilised lectins or glycans able to detect a wide range of analytes including whole cells/viruses are also discussed. PMID:27365034

  6. Lectins Identify Glycan Biomarkers on Glioblastoma-Derived Cancer Stem Cells

    PubMed Central

    Tucker-Burden, Carol; Chappa, Prasanthi; Krishnamoorthy, Malini; Gerwe, Brian A.; Scharer, Christopher D.; Heimburg-Molinaro, Jamie; Harris, Wayne; Usta, Sümeyra Naz; Eilertson, Carmen D.; Hadjipanayis, Constantinos G.; Stice, Steven L.; Brat, Daniel J.

    2012-01-01

    Glioblastoma (GBM) is a highly aggressive primary brain tumor with a poor prognosis. Despite aggressive therapy with surgery, radiotherapy, and chemotherapy, nearly all patients succumb to disease within 2 years. Several studies have supported the presence of stem-like cells in brain tumor cultures that are CD133-positive, are capable of self-renewal, and give rise to all cell types found within the tumor, potentially perpetuating growth. CD133 is a widely accepted marker for glioma-derived cancer stem cells; however, its reliability has been questioned, creating a need for other identifiers of this biologically important subpopulation. We used a panel of 20 lectins to identify differences in glycan expression found in the glycocalyx of undifferentiated glioma-derived stem cells and differentiated cells that arise from them. Fluorescently labeled lectins that specifically recognize α-N-acetylgalactosamine (GalNAc) and α-N-acetylglucosamine (GlcNAc) differentially bound to the cell surface based on the state of cellular differentiation. GalNAc and GlcNAc were highly expressed on the surface of undifferentiated cells and showed markedly reduced expression over a 12-day duration of differentiation. Additionally, the GalNAc-recognizing lectin Dolichos biflorus agglutinin was capable of specifically selecting and sorting glioma-derived stem cell populations from an unsorted tumor stock and this subpopulation had proliferative properties similar to CD133+ cells in vitro and also had tumor-forming capability in vivo. Our preliminary results on a single cerebellar GBM suggest that GalNAc and GlcNAc are novel biomarkers for identifying glioma-derived stem cells and can be used to isolate cancer stem cells from unsorted cell populations, thereby creating new cell lines for research or clinical testing. PMID:22435486

  7. Fungal lectins: a growing family.

    PubMed

    Kobayashi, Yuka; Kawagishi, Hirokazu

    2014-01-01

    Fungi are members of a large group of eukaryotic organisms that include yeasts and molds, as well as the most familiar member, mushrooms. Fungal lectins with unique specificity and structures have been discovered. In general, fungal lectins are classified into specific families based on their amino acid sequences and three-dimensional structures. In this chapter, we provide an overview of the approximately 80 types of mushroom and fungal lectins that have been isolated and studied to date. In particular, we have focused on ten fungal lectins (Agaricus bisporus, Agrocybe cylindracea, Aleuria aurantia, Aspergillus oryzae, Clitocybe nebularis, Marasmius oreades, Psathyrella velutina, Rhizopus stolonifer, Pholiota squarrosa, Polyporus squamosus), many of which are commercially available and their properties, sugar-binding specificities, structural grouping into families, and applications for biological research being described. The sialic acid-specific lectins (Agrocybe cylindracea and Polyporus squamosus) and fucose-specific lectins (Aleuria aurantia, Aspergillus oryzae, Rhizopus stolonifer, and Pholiota squarrosa) each showed potential for use in identifying sialic acid glycoconjugates and fucose glycoconjugates. Although not much is currently known about fungal lectins compared to animal and plant lectins, the knowledge accumulated thus far shows great promise for several applications in the fields of taxonomy, biomedicine, and molecular and cellular biology.

  8. Longitudinal Cerebral Fissure Anatomy Variations in Brachy-, Dolicho- and Mesaticephalic Dogs and Their Importance to Brain Surgery.

    PubMed

    Carreira, L Miguel; Ferreira, A

    2015-09-01

    The study used a sample of 69 formalin-fixed brains from adult dog cadavers (n = 69) and aimed (1) to characterize the longitudinal cerebral fissure (LCF) anatomy in brachy-(B), dolicho-(D) and mesaticephalic-(M) dogs and their potential differences, and (2) to establish cranioencephalic relationships between the LCF and five classical craniometric points(cp): asterion(ast), bregma(b), stephanion(st), glabella(g), and pterion(pt). Anatomical records were collected using a digital caliper, and for statistical analysis P-values < 0.05 were considered significant. The LCF length can be ranked, in ascending order as B < D < M, and if used as a surgical corridor, the M group had the greatest surgical corridor area, and the D group the smallest. LCF morphology was uniform among the three groups exhibiting dilated anterior(AR) and posterior(PR) regions and narrow middle region, where the most marked differences were registered. The LCF AR is the ideal spot to begin brain surgery if the LCF is to be used as a surgical corridor in B and M, while the LCF PR should be considered in D. The five cp selected were quite useful to understand LCF anatomical morphology, its projections over the external skull surface, and to establish cranioencephalic relationships between the LCF and vault; allowing us to consider the vault anterior area in B, and in M, and the vault posterior area in D for craniotomy initial burr-hole placement. As for the distance from the cp projections in the brain surface to the LCF regions, major differences were registered by comparing the B group with the other two, and for both hemispheres. © 2015 Wiley Periodicals, Inc.

  9. The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study.

    PubMed

    Jones, Carolyn J P; Wilsher, Sandra A; Wooding, F B P; Benirschke, K; Allen, W R

    2015-02-01

    The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. Some differences between Okapi and Giraffe were evident. Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development.

  10. Affinity cytochemistry of vascular endothelia in brain tumors by biotinylated Ulex europaeus type I lectin (UEA I).

    PubMed

    Weber, T; Seitz, R J; Liebert, U G; Gallasch, E; Wechsler, W

    1985-01-01

    The vascularization of 50 tumors of the central nervous system (CNS) including 17 meningiomas, 25 neuroectodermal tumors, i.e., astrocytomas, oligodendrogliomas, mixed gliomas, glioblastomas, medulloblastomas, seven metastatic carcinomas, and one malignant hemangioendothelioma were investigated using biotinylated Ulex europaeus type I lectin (UEA I) in an indirect avidinbiotin-peroxidase procedure. The cytochemical staining pattern of UEA I on paraffin sections was compared with that of biotinylated Dolichos biflorus lectin (DBA), and with the immunocytochemical staining of factor VIII related antigen (F VIII/RAG) by polyclonal antisera using the PAP technique. UEA I visualized the endothelia of blood vessels with equal intensity, sensitivity, and reliability in normal brain and in tumor tissue with neovascularization. While large, medium, and small vessels were equally well demonstrated by UEA I and antibodies against FVIII/RAG, capillaries and endothelial sprouts were stained more consistently and intensely by UEA I. No reliable cytochemical staining could be obtained by DBA regardless of tissue or cell type investigated. It is concluded that UEA I is a highly useful cytochemical marker for the identification of vascular endothelia in paraffin sections of human brain tumors.

  11. Lectins and Tetrahymena - A review.

    PubMed

    Csaba, György

    2016-09-01

    The unicellular ciliate Tetrahymena is a complete organism, one of the most highly developed protozoans, which has specialized organelles performing each of the functions characteristic to the cells of higher ranked animals. It is also able to produce, store, and secrete hormones of higher ranked animals and also react to them. It produces lectins that can bind them and has functions, which are influenced by exogenous lectins. The review lists the observations on the relationship between lectins and Tetrahymena and try to construe them on the basis of the data, which are at our disposal. Considering the data, lectins can be used by Tetrahymena as materials for influencing conjugation, for stimulating hormone receptors, and by this, mimic the hormonal functions. Lectins can influence phagocytosis and movement of the cells as well as the cell division. As Tetrahymena can recognize both related and hostile cells by the help of lectins and surface sugars, it could be surmised a complex predator-prey system. This could determine the survival of the population as well as the nourishment conditions. When Tetrahymena is pathogenic, it can use lectins as virulence factors.

  12. Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans.

    PubMed

    Aritua, Valente; Harrison, James; Sapp, Melanie; Buruchara, Robin; Smith, Julian; Studholme, David J

    2015-01-01

    Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analyzed the genomes of 26 strains of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial strain from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type strain (NCPPB 3035). The strains from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans.

  13. Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans

    PubMed Central

    Aritua, Valente; Harrison, James; Sapp, Melanie; Buruchara, Robin; Smith, Julian; Studholme, David J.

    2015-01-01

    Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analyzed the genomes of 26 strains of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial strain from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type strain (NCPPB 3035). The strains from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans. PMID:26500625

  14. Evaluation of maize yield in an on-farm maize-soybean and maize-Lablab crop rotation systems in the Northern Guinea Savanna of Nigeria.

    PubMed

    Okogun, J A; Sanginga, N; Abaidoo, R C

    2007-11-01

    An attempt was made to solving the problem of shortfall of fertilizer to maize production in the Northern Guinea Savanna (NGS) of Nigeria by harnessing the potentials of legume/cereal crop rotation in on-farm trials. The yield of maize that succeeded two soybean varieties and Lablab in a two-cycle of soybean/maize and Lablab/maize crop rotation in NGS Nigeria was assessed in researcher-managed and farmer-managed plots. Though maize that followed the soybean received between 5 kg N ha(-1) from improved soybean variety (TGx 1448-2E) and 17 kg N ha(-1) from farmer soybean variety (Samsoy-2) as N balance, this did not significantly (p = 0.05) affect the maize yields. The soybean shed 90-100% of its leaves at physiological maturity which resulted in about 110 kg N ha(-1) N uptake. This source of N might be one of the factors responsible for the increase in maize yield that followed soybean (20 to 24%) compared with continuous maize yield plot. Maize yield in previous Lablab plot was significantly (p = 0.05) higher than in all other treatments. Maize yield in farmer-managed plot ranged between 0.13 and 4.53 t ha(-1), maize yield in researcher-managed plot was over 200% higher than maize yield in farmer-managed plot because of poor crop management on the part of the farmer.

  15. Use of lectins in immunohematology

    PubMed Central

    Gorakshakar, Ajit C.; Ghosh, Kanjaksha

    2016-01-01

    Lectins are carbohydrate binding proteins present in seeds of many plants, especially corals and beans, in fungi and bacteria, and in animals. Apart from their hemagglutinating property, a wide range of functions have been attributed to them. Their importance in the area of immunohematology is immense. They are used to detect specific red cell antigens, to activate different types of lymphocytes, in order to resolve problems related to polyagglutination and so on. The introduction of advanced biotechnological tools generates new opportunities to exploit the properties of lectins, which were not used earlier. Stem cell research is a very important area in transplant medicine. Certain lectins detect surface markers of stem cell. Hence, they are used to understand the developmental biology of stem cells. The role of various lectins in the areas of transfusion and transplant medicine is discussed in detail in this review. PMID:27011665

  16. A review of fish lectins.

    PubMed

    Ng, Tzi Bun; Fai Cheung, Randy Chi; Wing Ng, Charlene Cheuk; Fang, Evandro Fei; Wong, Jack Ho

    2015-01-01

    Lectins have been reported from various tissues of a diversity of fish species including Japanese eel, conger eel, electric eel, bighead carp, gibel carp, grass carp, Arabian Gulf catfish, channel catfish, blue catfish, catfish, pike perch, perch, powan, zebrafish, toxic moray, cobia fish, steelhead trout, Japanese trout, Atlantic salmon, chinook salmon, olive rainbow smelt, rainbow smelt, white-spotted charr, tilapia, blue gourami, ayu, Potca fish, Spanish mackerel, gilt head bream, tench, roach, rudd, common skate, and sea lamprey. The tissues from which the lectins were isolated comprise gills, eggs, electric organ, stomach, intestine, and liver. Lectins have also been isolated from skin, mucus serum, and plasma. The lectins differ in molecular weight, number of subunits, glycosylation, sugar binding specificity and amino acid sequence. Their activities include antimicrobial, antitumor, immunoregulatory and a role in development.

  17. Lectins in the investigation of receptors

    NASA Astrophysics Data System (ADS)

    Lakhtin, V. M.; Yamskov, Igor A.

    1991-08-01

    Problems of the purification and characterisation are considered for approximately 270 receptors (including cell surface and organelle enzymes), which are glycoconjugates (mainly glycoproteins) from animals, plants and microorganisms, using various lectins (mainly lectin sorbents). An analysis has been carried out of the stages of lectin affinity chromatography of receptors (choice of detergent, use of organic solvents, elution with carbohydrates, etc.). Examples are given of procedures for the purification of receptors, including the use of paired columns and combination chromatography on lectins. The possibility of separating sub-populations of receptors using lectins has been demonstrated. Examples are given of the use of lectins in the analysis of the oligosaccharide structure of receptors. Cases are recorded of the interaction of receptors with endogenous lectins and of receptor lectins with endogenous glycoconjugates. It has been shown that lectins, in combination with glycosidases and antibodies, may be useful in the investigation of receptors. The bibliography contains 406 references.

  18. Lectins in human pathogenic fungi.

    PubMed

    Gallegos, Belém; Martínez, Ruth; Pérez, Laura; Del Socorro Pina, María; Perez, Eduardo; Hernández, Pedro

    2014-01-01

    Lectins are carbohydrate-binding proteins widely distributed in nature. They constitute a highly diverse group of proteins consisting of many different protein families that are, in general, structurally unrelated. In the last few years, mushroom and other fungal lectins have attracted wide attention due to their antitumour, antiproliferative and immunomodulatory activities. The present mini-review provides concise information about recent developments in understanding lectins from human pathogenic fungi. A bibliographic search was performed in the Science Direct and PubMed databases, using the following keywords "lectin", "fungi", "human" and "pathogenic". Lectins present in fungi have been classified; however, the role played by lectins derived from human pathogenic fungi in infectious processes remains uncertain; thus, this is a scientific field requiring more research. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012). Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  19. Persistent torticollis, facial asymmetry, grooved tongue, and dolicho-odontoid process in connection with atlas malformation complex in three family subjects

    PubMed Central

    Chehida, Farid Ben; Gharbi, Hassan; Ghachem, Maher Ben; Grill, Franz; Varga, Franz; Klaushofer, Klaus

    2007-01-01

    Congenital clefts and other malformations of the atlas are incidental findings identified while investigating the cervical spine following trauma. A persistent bifid anterior and posterior arch of the atlas beyond the age of 3–4 years is observed in skeletal dysplasias, Goldenhar syndrome, Conradi syndrome, and Down’s syndrome. There is a high incidence of both anterior and posterior spina bifida of the atlas in patients with metabolic disorders, such as Morquio’s syndrome [Baraitser and Winter in London dysmorphology database, Oxford University Press, 2005; Torriani, Lourenco in Rev Hosp Clin Fac Med Sao Paulo 53: 73–76, 2002]. We report two siblings and their mother, with congenital, persistent torticollis, plagiocephaly, facial asymmetry, grooved tongues, and asymptomatic “dolicho-odontoid process”. All are of normal intelligence. No associated Neurological dysfunction, paresis, apnoea, or failures to thrive were encountered. Radiographs of the cervical spine were non-contributory, but 3D CT scanning of this area allowed further visualisation of the cervico–cranial malformation complex in this family and might possibly explain the sudden early juvenile mortality. Agenesis of the posterior arch of the atlas and bifidity/clefting of anterior arch of the atlas associated with asymptomatic “dolicho-odontoid process” were the hallmark in the proband and his female sibling. Some of the features were present in the mother. All the family subjects were investigated. To the best of our knowledge the constellation of malformation complex in this family has not been previously reported. PMID:17245565

  20. Development of lectin-linked immunomagnetic separation for the detection of hepatitis a virus.

    PubMed

    Ko, Sang-Mu; Kwon, Joseph; Vaidya, Bipin; Choi, Jong Soon; Lee, Hee-Min; Oh, Myung-Joo; Bae, Hyeun-Jong; Cho, Se-Young; Oh, Kyung-Seo; Kim, Duwoon

    2014-03-04

    The accuracy and sensitivity of PCR-based methods for detection of hepatitis A virus (HAV) are dependent on the methods used to separate and concentrate the HAV from the infected cells. The pH and ionic strength affect the binding affinity of the virus to cells. In this study, we initially investigated the effects of pH (4.0-10.0) and metal ions (Fe²⁺, Co²⁺, Cu²⁺, Mg²⁺, K⁺, and Ca²⁺) on the binding of HAV to oyster digestive cells. The lowest relative binding (RB) of HAV to the cells was found at pH 4.0 and in FeSO₄ solution (64.6% and 68.1%, respectively). To develop an alternative to antibody-dependent immunomagnetic separation prior to detection of HAV using RT-PCR, the binding of HAV to five lectins, peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Ulex europaeus agglutinin (UEA-1) and soybean agglutinin (SBA), was evaluated using ELISAs. SBA showed significantly higher RB to HAV than the other lectins tested. In addition, HAV could be concentrated within 30 min using SBA-linked magnetic bead separation (SMS) prior to the RT-PCR assay. Our findings demonstrate the feasibility of using SMS combined with RT-PCR to detect HAV at dilutions ranging from 10⁻¹-10⁻⁴ of a HAV stock (titer: 10⁴ TCID₅₀/mL).

  1. Lectin-Like Constituents of Foods Which React with Components of Serum, Saliva, and Streptococcus mutans

    PubMed Central

    Gibbons, R. J.; Dankers, I.

    1981-01-01

    Hot and cold aqueous extracts were prepared from 22 commonly ingested fruits, vegetables, and seeds. When tested by agar diffusion, extracts from 13 and 10 of the foods formed precipitin bands with samples of normal rabbit serum and human saliva, respectively; extracts from four of the foods also reacted with antigen extracts of strains of Streptococcus mutans. When added to rabbit antiserum, extracts from 18 of 21 foods tested inhibited reactivity with antigen extracts derived from S. mutans MT3. Extracts from 16 foods agglutinated whole S. mutans cells, whereas those from 10 foods agglutinated human erythrocytes of blood types A and B. The lectin-like activities of extracts which reacted with human saliva were studied further. Pretreatment of saliva-coated hydroxyapatite (S-HA) beads with extracts of bananas, coconuts, carrots, alfalfa, and sunflower seeds markedly reduced the subsequent adsorption of S. mutans MT3. Pretreatment of S-HA with banana extract also strongly inhibited adsorption of S. mutans H12 and S. sanguis C1, but it had little effect on attachment of Actinomyces naeslundii L13 or A. viscosus LY7. Absorption experiments indicated that the component(s) in banana extract responsible for inhibiting streptococcal adsorption to S-HA was identical to that which bound to human erythrocytes. The banana hemagglutinin exhibited highest activity between pH 7 and 8, and it was inhibited by high concentrations of glucosamine, galactosamine, and, to a lesser extent, mannosamine. Other sugars tested had no effect. The selective bacterial adsorption-inhibiting effect noted for banana extract was also observed in studies with purified lectins. Thus, pretreating S-HA with wheat germ agglutinin and concanavalin A inhibited adsorption of S. mutans MT3 cells, whereas peanut agglutinin, Ulex agglutinin, Dolichos agglutinin, and soybean agglutinin had little effect; none of these lectins affected attachment of A. viscosus LY7. Collectively, the observations suggest that

  2. Dietary Plant Lectins Appear to Be Transported from the Gut to Gain Access to and Alter Dopaminergic Neurons of Caenorhabditis elegans, a Potential Etiology of Parkinson’s Disease

    PubMed Central

    Zheng, Jolene; Wang, Mingming; Wei, Wenqian; Keller, Jeffrey N.; Adhikari, Binita; King, Jason F.; King, Michael L.; Peng, Nan; Laine, Roger A.

    2016-01-01

    Lectins from dietary plants have been shown to enhance drug absorption in the gastrointestinal tract of rats, be transported trans-synaptically as shown by tracing of axonal and dendritic paths, and enhance gene delivery. Other carbohydrate-binding protein toxins are known to traverse the gut intact in dogs. Post-feeding rhodamine- or TRITC-tagged dietary lectins, the lectins were tracked from gut to dopaminergic neurons (DAergic-N) in transgenic Caenorhabditis elegans (C. elegans) [egIs1(Pdat-1:GFP)] where the mutant has the green fluorescent protein (GFP) gene fused to a dopamine transport protein gene labeling DAergic-N. The lectins were supplemented along with the food organism Escherichia coli (OP50). Among nine tested rhodamine/TRITC-tagged lectins, four, including Phaseolus vulgaris erythroagglutinin (PHA-E), Bandeiraea simplicifolia (BS-I), Dolichos biflorus agglutinin (DBA), and Arachis hypogaea agglutinin (PNA), appeared to be transported from gut to the GFP-DAergic-N. Griffonia Simplicifolia and PHA-E, reduced the number of GFP-DAergic-N, suggesting a toxic activity. PHA-E, BS-I, Pisum sativum (PSA), and Triticum vulgaris agglutinin (Succinylated) reduced fluorescent intensity of GFP-DAergic-N. PHA-E, PSA, Concanavalin A, and Triticum vulgaris agglutinin decreased the size of GFP-DAergic-N, while BS-I increased neuron size. These observations suggest that dietary plant lectins are transported to and affect DAergic-N in C. elegans, which support Braak and Hawkes’ hypothesis, suggesting one alternate potential dietary etiology of Parkinson’s disease (PD). A recent Danish study showed that vagotomy resulted in 40% lower incidence of PD over 20 years. Differences in inherited sugar structures of gut and neuronal cell surfaces may make some individuals more susceptible in this conceptual disease etiology model. PMID:27014695

  3. Lectin-probed western blot analysis.

    PubMed

    Sato, Takeshi

    2014-01-01

    Lectin-probed western blot analysis, the so-called lectin blot analysis, is a useful method to yield basic information on the glycan structures of glycoproteins, based on the carbohydrate-binding specificities of lectins. By lectin blot analysis, researchers can directly analyze the glycan structures without releasing the glycans from glycoproteins. Here, the author describes protocols for standard analysis, and applies analysis in combination with glycosidase digestion of blot.

  4. Monitoring lectin interactions with carbohydrates.

    PubMed

    de Bentzmann, Sophie; Varrot, Annabelle; Imberty, Anne

    2014-01-01

    Protein-carbohydrate interactions are often involved in the first step of infection and Pseudomonas aeruginosa produces several proteins that are able to bind specifically to glycan epitopes present on host epithelia. The experimental approaches for studying protein-carbohydrate interaction have been inspired, with some adaptations, from those commonly used for protein-protein or protein-ligand interactions. A range of methods are described herein for detecting lectin activity, screening for monosaccharide or oligosaccharide specificity, determining the affinity of binding together with thermodynamics and kinetics parameters, and producing crystal of lectin-carbohydrate complexes for further structural studies.

  5. Effects of an oxidative agent and lectins on the binding inhibition of recombinant hepatitis a virus proteins to oyster digestive tissues.

    PubMed

    Ko, Sang-Mu; Kim, Jong-Oh; Oh, Myung-Joo; Kim, Duwoon

    2011-01-01

    While the exact mechanism of hepatitis A virus (HAV) accumulation remains unclear, it has been demonstrated that viruses related to shellfish-borne gastroenteritis can bind to carbohydrates of oyster tissues. We investigated carbohydrate-binding sites to determine if they were related to the binding of HAV to carbohydrate moieties on oyster digestive tissues (DTs) using recombinant HAV proteins (rHAVPs). In addition, we evaluated lectins to determine if they influenced the inhibition of binding of rHAVPs to carbohydrates present in DT. DT that was treated with 0.5% potassium periodate allowed only 23% ± 3.6% and 33% ± 7.8% binding of VP1-P2A and VP1 rHAVPs, respectively, when compared with a control group (100%) treated with distilled water, indicating that carbohydrate-binding sites are strongly related to the binding of HAV. Soybean agglutinin (SBA) led to the greatest decrease in the binding affinity among six lectins (Helix pomatia, Dolichos biflorus, Ulex europaeus, SBA, Triticum vulgaris, and Arachis hypogaea) tested for inhibition of the binding of rHAVPs to DT, indicating that exposing the virus-contaminated DT to SBA might have the potential to depurate viral contaminants found in shellfish food products by high-affinity binding between SBA and rHAVPs, thus improving food safety.

  6. An overview of lectins purification strategies.

    PubMed

    Nascimento, Kelany S; Cunha, Ana I; Nascimento, Kyria S; Cavada, Benildo S; Azevedo, Ana M; Aires-Barros, Maria Raquel

    2012-11-01

    Lectins hold great promise not only as reagents for diagnostics and drug discovery but also as a novel class of biopharmaceutical products. In fact, new research directions in the last years have led to major developments in the uses of plant lectins as therapeutic agents against numerous diseases in an ageing society. It is even expected that lectins may occupy an important place in the biopharmaceutical industry next to monoclonal antibodies. All these new trends are placing a tremendous emphasis on the development of new approaches for faster lectins development, selection, and optimization, including alternatives methods of purification. This article reviews the isolation and purification methods used for lectins purification. Origins and applications of lectins are described, highlighting the special features of this class of proteins, such as the carbohydrated-binding domains and their importance in the development of affinity methodologies to increase and facilitate lectins purification. Published strategies for the purification of lectins from different sources are analyzed in relation to the purification methods used, their sequence, and the number of times they are used in a purification procedure. The purity of lectins is analyzed in relation to the average overall yield and purification factors obtained for each purification scheme for these proteins and the purification steps necessary. New directions are described for improving lectins separation and purification. Copyright © 2012 John Wiley & Sons, Ltd.

  7. Effect of intercropping Panicum maximum var. Ntchisi and Lablab purpureus on the growth, herbage yield and chemical composition of Panicum maximum var. Ntchisi at different harvesting times.

    PubMed

    Ojo, V O A; Dele, P A; Amole, T A; Anele, U Y; Adeoye, S A; Hassan, O A; Olanite, J A; Idowu, O J

    2013-11-15

    The study was conducted to evaluate the effect of intercropping Panicum maximum var. Ntchisi and Lablab purpureus on the growth, herbage yield and chemical composition of P. maximum var. Ntchisi at different harvesting times at the Teaching and Research farm, Federal University of Agriculture, Abeokuta in a randomized complete block design. Samples were collected at different harvesting times (8, 10, 12, 14 weeks after planting). The growth parameters which were plant height, leaf length, leaf number and tiller number measured showed that the intercropping of grass with legume were higher than in the sole plot of P. maximum var. Ntchisi. The plant yield was consistently higher (p < 0.05) in intercropped forages than in sole throughout the harvesting times. The crude protein contents of the forages were also higher for the intercropped across the treatments. The values of the fibre components were significantly different (p < 0.05) at different harvesting times and it was increasing as the harvesting time was increasing. From this study, considering the herbage yield and chemical composition of intecropping Panicum maximum var. Ntchisi and Lablab purpureus, they can be grazed by ruminant animals or harvested at 12 weeks after planting when the quality and quantity will support livestock productivity and can be conserved to be fed to ruminant animals during dry season when feed availability and quality are extremely low.

  8. Lectin cDNA and transgenic plants derived therefrom

    DOEpatents

    Raikhel, Natasha V.

    2000-10-03

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

  9. Microbial lectins and their prospective mitogenic potential.

    PubMed

    Singh, Ram Sarup; Walia, Amandeep K

    2014-11-01

    The binding of mitogenic lectins to the glycoconjugates on cell surface receptor triggers multitude of reactions involving different signal transduction pathways which ultimately results in cell proliferation. Since 1960 after the chance discovery of mitogenic property of lectins by Nowell, it has attracted more attention. The property of stimulation of mature lymphocytes aided the immunological investigations to study and analyse the mechanism of cell stimulation and division. Plant lectins are extensively studied for mitogenic activity and their widespread applications are also reported. During the past two decades, the study of biological properties of microbial lectins has increased tremendously. Their biological activities including mitogenic potential are equally applicable as that of plant lectins. The review mainly focuses on mitogenic potential of microbial lectins. It will provide a comprehensive view regarding distribution of this remarkable property among microbes including algae, fungi and bacteria, mechanisms and models of mitogenic stimulation, and also assays being employed to detect the mitogenic activity.

  10. Plant as a plenteous reserve of lectin

    PubMed Central

    Hivrale, AU; Ingale, AG

    2013-01-01

    Lectins are clusters of glycoproteins of nonimmune foundation that combine specifically and reversibly to carbohydrates, mainly the sugar moiety of glycoconjugates, resulting in cell agglutination and precipitation of glycoconjugates. They are universally distributed in nature, being established in plants, fungi, viruses, bacteria, crustacea, insects, and animals, but leguminacae plants are rich source of lectins. The present review reveals the structure, biological properties, and application of plant lectins. PMID:24084524

  11. SUEL-related lectins, a lectin family widely distributed throughout organisms.

    PubMed

    Tateno, Hiroaki

    2010-01-01

    Glycan-binding proteins are categorized into two groups, lectins and sulfated glycosaminoglycan-binding proteins. SUEL-related lectins are members of a superfamily of proteins containing a carbohydrate-recognition domain (CRD), which is structurally similar to sea urchin egg lectin (SUEL). Here I review the structure and function of this family of proteins.

  12. Influence of Lectins on Constricting Ring Formation by Arthrobotrys dactyloides.

    PubMed

    Kaplan, D T; Davis, E L; Walter, D E

    1991-04-01

    Incubation of Arthrobotrys dactyloides conidia in the presence of Radopholus citrophilus in lectin solutions with their corresponding sugars did not alter the stimulation of trap formation in solutions containing lectins alone. The lack of inhibition of lectin-stimulated trap formation by sugars or by lectin denaturation and the lack of lectin specificity indicate that the carbohydrate-binding regions of the particular lectins studied are not the stimulatory moieties of these macromolecules.

  13. Lectins and their application to clinical microbiology.

    PubMed Central

    Slifkin, M; Doyle, R J

    1990-01-01

    Lectins are generally associated with plant or animal components, selectively bind carbohydrates, and interact with procaryotic and eucaryotic cells. Lectins have various specificities that are associated with their ability to interact with acetylaminocarbohydrates, aminocarbohydrates, sialic acids, hexoses, pentoses, and as other carbohydrates. Microbial surfaces generally contain many of the sugar residues that react with lectins. Lectins are presently used in the clinical laboratory to type blood cells and are used in a wide spectrum of applications, including, in part, as carriers of chemotherapeutic agents, as mitogens, for fractionation of animal cells, and for investigations of cellular surfaces. Numerous studies have shown that lectins can be used to identify rapidly certain microorganisms isolated from a clinical specimen or directly in a clinical specimen. Lectins have been demonstrated to be important diagnostic reagents in the major realms of clinical microbiology. Thus, they have been applied in bacteriology, mycology, mycobacteriology, and virology for the identification and/or differentiation of various microorganisms. Lectins have been used successfully as epidemiologic as well as taxonomic markers of specific microorganisms. Lectins provide the clinical microbiologist with cost-effective and potential diagnostic reagents. This review describes the applications of lectins in clinical microbiology. Images PMID:2200603

  14. Fungal lectins: structure, function and potential applications.

    PubMed

    Varrot, Annabelle; Basheer, Soorej M; Imberty, Anne

    2013-10-01

    Lectins are a widespread class of proteins implicated in many essential cellular and molecular recognition processes. They recognize carbohydrates in a non-catalytic, specific and reversible manner. Fungi, which include mushrooms, microfungi and yeasts, have attracted wide interest in recent years. They are indeed a promising source for novel lectins with unique specificity and potential for biomedical and biotechnological applications. Information on fungal lectins, particularly structural insight, is scarce compared to that on their plant and animal counterparts. This review therefore focuses on the structure, function, and exploitable properties of fungal lectins. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Perspectives in glycomics and lectin engineering.

    PubMed

    Tkac, Jan; Bertok, Tomas; Nahalka, Jozef; Gemeiner, Peter

    2014-01-01

    This chapter would like to provide a short survey of the most promising concepts applied recently in analysis of glycoproteins based on lectins. The first part describes the most exciting analytical approaches used in the field of glycoprofiling based on integration of nanoparticles, nanowires, nanotubes, or nanochannels or using novel transducing platforms allowing to detect very low levels of glycoproteins in a label-free mode of operation. The second part describes application of recombinant lectins containing several tags applied for oriented and ordered immobilization of lectins. Besides already established concepts of glycoprofiling several novel aspects, which we think will be taken into account for future, more robust glycan analysis, are described including modified lectins, peptide lectin aptamers, and DNA aptamers with lectin-like specificity introduced by modified nucleotides. The last part of the chapter describes a novel concept of a glycocodon, which can lead to a better understanding of glycan-lectin interaction and for design of novel lectins with unknown specificities and/or better affinities toward glycan target or for rational design of peptide lectin aptamers or DNA aptamers.

  16. Marine lectins and their medicinal applications.

    PubMed

    Cheung, Randy Chi Fai; Wong, Jack Ho; Pan, Wenliang; Chan, Yau Sang; Yin, Cuiming; Dan, Xiuli; Ng, Tzi Bun

    2015-05-01

    Marine organisms have been extensively explored for the last several decades as potential sources of novel biologically active compounds, and extensive research has been conducted on lectins. Lectins derived from marine organisms are structurally diverse and also differ from those identified from terrestrial organisms. Marine lectins appear to be particularly useful in some biological applications. They seem to induce negligible immunogenicity because they have a relatively small size, are more stable due to extensive disulfide bridge formation, and have high specificity for complex glyco-conjugates and carbohydrates instead of simple sugars. It is clear that many of them have not yet been extensively studied when compared with their terrestrial counterparts. Marine lectins can be used to design and develop new potentially useful therapeutic agents. This review encompasses recent research on the isolation and identification of marine lectins with potential value in medicinal applications.

  17. Lectin microarray technology identifies specific lectins related to lymph node metastasis of advanced gastric cancer.

    PubMed

    Yamashita, Keishi; Kuno, Atsushi; Matsuda, Atsushi; Ikehata, Yuzuru; Katada, Natsuya; Hirabayashi, Jun; Narimatsu, Hisashi; Watanabe, Masahiko

    2016-04-01

    Although various molecular profiling technologies have the potential to predict specific tumor phenotypes, the comprehensive profiling of lectin-bound glycans in human cancer tissues has not yet been achieved. We examined 242 advanced gastric cancer (AGC) patients without or with lymph node metastasis-N0 (n = 62) or N+ (n = 180)-by lectin microarray, and identified the specific lectins highly associated with AGC phenotypes. In seven gastric cancer cell lines, in contrast to expressed-in-cancer lectins, not-expressed-in-cancer (NEC) lectins were tentatively designated by lectin microarray. Binding signals of the specific lectins were robustly reduced in AGC patients with N+ status as compared with those with N0 status. The receiver operating characteristic curve determined the optimal cutoff value to differentiate N0 status from N+ status, and subsequent profiling of NEC lectins identified Vicia villosa agglutinin (VVA) association with the significant other lectins involved in lymph node metastasis. VVA reaction was clearly found on cancer cells, suggesting that it may result from carcinoma-stroma interaction in primary AGC, because VVA is an NEC lectin. Most intriguingly, VVA reaction was remarkably attenuated in the tumor cells of the metastatic lymph nodes, even if it was recognized in primary AGC. In AGC, histological type was strongly associated with soybean agglutinin and Bauhinia purpurea lectin, whereas p53 mutation was the best correlated with Griffonia simplicifolia lectin II. Lectin microarrays can be used to very accurately quantify the reaction of glycans with tumor tissues, and such profiles may represent the specific phenotypes, including N+ status, histological type, or p53 mutation of AGC.

  18. A new variant of antimetabolic protein, arcelin from an Indian bean, Lablab purpureus (Linn.) and its effect on the stored product pest, Callosobruchus maculatus.

    PubMed

    Janarthanan, Sundaram; Sakthivelkumar, Shanmugavel; Veeramani, Velayutham; Radhika, Dixit; Muthukrishanan, Subbaratnam

    2012-12-15

    The anti-metabolic or insecticidal gene, arcelin (Arl) was isolated, cloned and sequenced using sequence specific degenerate primers from the seeds of Lablab purpureus collected from the Western Ghats, Tamil Nadu, India. The L. purpureus arcelin nucleotide sequence was homologous to Arl-3 and Arl-4 alleles from Phaseolus spp. The protein it encodes has 70% amino acid identity with the amino acid sequences of Arl-3I, Arl-3III, Arl-4 precursor, Arl-4 and Arl-4I. The partially purified arcelin from the seeds of L. purpureus using an artificial diet confirmed the complete retardation of development of the stored product pest Callosobruchus maculatus at 0.2% w/w arcelin-incorporated artificial seeds.

  19. Overall strategy for functional analysis of animal lectins.

    PubMed

    Kawasaki, Norihito

    2014-01-01

    Animal lectins elicit biological functions through the interaction with glycan ligands. To clarify the functions of the lectins, both identification of their glycan ligand structure and assessment of impact of lectin-glycan interaction on the biological event are essential strategies. This chapter focuses on two of key useful methodologies for planning experiments based on the strategies. One is the detection of lectin-glycan interaction by the multivalent display of lectins and glycans. This methodology is a powerful means for identification of the glycan ligand structure and proteins and/or lipids carrying the glycan ligands for lectins. The other is the intervention of lectin-glycan interaction to assess the biological roles of lectins. Bioinformatics especially useful for animal lectins will be also described in this chapter. The concepts described in this chapter are versatile and applicable to a wide range of animal lectin research.

  20. Lectins in human cancer: both a devil and an angel?

    PubMed

    Dan, Xiu Li; Ng, Tzi Bun

    2013-09-01

    Lectins are a group of proteins which could recognize different sugar structures and specifically initiate reversible binding with them. Lectins are universally expressed in different organisms and undertake important biological roles. Understanding of their inherent roles and mechanisms that they employ has inspired researches with new ideas and applications. For example, along with the revelation of their anti-insect function, plant lectins exhibit great potential in agriculture. In human beings, lectins shoulder great missions in cell communication, differentiation and vesicle trafficking etc., aberrant expression of lectins is always associated with diseases. Mannan-binding lectin deficiency leads to immune disorder and liver sinusoidal endothelial cell lectin is involved in colorectal carcinoma liver metastasis. In this review, we present two contradictory roles of lectins in human cancer: the promotive roles of homologous lectins and suppressive roles of heterologous lectins in cancer development. Hopefully, this review will facilitate a better understanding of tumorigenesis and provide references for cancer treatment.

  1. C-type lectins facilitate tumor metastasis

    PubMed Central

    Ding, Dongbing; Yao, Yao; Zhang, Songbai; Su, Chunjie; Zhang, Yonglian

    2017-01-01

    Metastasis, a life-threatening complication of cancer, leads to the majority of cases of cancer-associated mortality. Unfortunately, the underlying molecular and cellular mechanisms of cancer metastasis remain to be fully elucidated. C-type lectins are a large group of proteins, which share structurally homologous carbohydrate-recognition domains (CRDs) and possess diverse physiological functions, including inflammation and antimicrobial immunity. Accumulating evidence has demonstrated the contribution of C-type lectins in different steps of the metastatic spread of cancer. Notably, a substantial proportion of C-type lectins, including selectins, mannose receptor (MR) and liver and lymph node sinusoidal endothelial cell C-type lectin, are important molecular targets for the formation of metastases in vitro and in vivo. The present review summarizes what has been found regarding C-type lectins in the lymphatic and hematogenous metastasis of cancer. An improved understanding the role of C-type lectins in cancer metastasis provides a comprehensive perspective for further clarifying the molecular mechanisms of cancer metastasis and supports the development of novel C-type lectins-based therapies the for prevention of metastasis in certain types of cancer. PMID:28123516

  2. Mushroom Lectins as Promising Anticancer Substances.

    PubMed

    Singh, Ram Sarup; Kaur, Hemant Preet; Kanwar, Jagat Rakesh

    2016-01-01

    Lectins are proteins/glycoproteins of non-immune origin, which are widely distributed in nature. They have at least one non-catalytic domain, which binds reversibly to specific monosaccharides or oligosaccharides. Lectins recognizing sugar moieties in cell walls or cell membranes alter the membrane physiology and trigger biochemical changes in the cell. Thus, various applications of lectins have been described, for example as tools to identify aberrant glycans expressed by neoplastic cells and as antitumor agents by inducing apoptosis by various mechanisms. In order to widen applications of anti-tumor lectins, a detailed investigation of their action mechanism is required. Mushrooms are a valuable source of novel lectins with unique specificities and potentials for biotechnological and biomedical applications. This article reviews information on anti-proliferative activity of mushroom lectins obtained in-vitro and in-vivo. The possible role of lectins as cancer therapeutics is discussed together with the mechanisms underlying the anti-proliferative activity, which may help to exploit these biomolecules as potential novel antitumor drugs in near future.

  3. C-type lectins facilitate tumor metastasis.

    PubMed

    Ding, Dongbing; Yao, Yao; Zhang, Songbai; Su, Chunjie; Zhang, Yonglian

    2017-01-01

    Metastasis, a life-threatening complication of cancer, leads to the majority of cases of cancer-associated mortality. Unfortunately, the underlying molecular and cellular mechanisms of cancer metastasis remain to be fully elucidated. C-type lectins are a large group of proteins, which share structurally homologous carbohydrate-recognition domains (CRDs) and possess diverse physiological functions, including inflammation and antimicrobial immunity. Accumulating evidence has demonstrated the contribution of C-type lectins in different steps of the metastatic spread of cancer. Notably, a substantial proportion of C-type lectins, including selectins, mannose receptor (MR) and liver and lymph node sinusoidal endothelial cell C-type lectin, are important molecular targets for the formation of metastases in vitro and in vivo. The present review summarizes what has been found regarding C-type lectins in the lymphatic and hematogenous metastasis of cancer. An improved understanding the role of C-type lectins in cancer metastasis provides a comprehensive perspective for further clarifying the molecular mechanisms of cancer metastasis and supports the development of novel C-type lectins-based therapies the for prevention of metastasis in certain types of cancer.

  4. Agglutination of Helicobacter pylori coccoids by lectins

    PubMed Central

    Khin, Mar Mar; Hua, Jie Song; Ng, Han Cong; Wadström, Torkel; Ho, Bow

    2000-01-01

    AIM: To study the agglutination pattern of Helicobacter pylori coccoid and spiral forms. METHODS: Assays of agglutination and agglutination inhibition were applied using fifteen commercial lectins. RESULTS: Strong agglutination was observed with mannose-specific Concanavalin A (Con A), fucose-specific Tetragonolobus purpureas (Lotus A) and N-acetyl glucosamine-specific Triticum vulgaris (WGA) lectins. Mannose and fucose specific lectins were reactive with all strains of H. pylori coccoids as compared to the spirals. Specific carbohydrates, glycoproteins and mucin were shown to inhibit H. pylori lectin-agglutination reactions. Pre-treatment of the bacterial cells with formalin and sulphuric acid did not alter the agglutination patterns with lectins. However, sodium periodate treatment of bacterial cells were shown to inhibit agglutination reaction with Con A, Lotus A and WGA lectins. On the contrary, enzymatic treatment of coccoids and spirals did not show marked inhibition of H. pylori lectin agglutination. Interes tingly, heating of H. pylori cells at 60 °C for 1 h was shown to augment the agglutination with all of the lectins tested. CONCLUSION: The considerable differences in lectin agglutination patterns seen among the two differentiated forms of H. pylori might be attributable to the structural changes during the events of morphological transformation, resulting in exposing or masking some of the sugar residues on the cell surface. Possibility of various sugar residues on the cell wall of the coccoids may allow them to bind to different carbohydrate receptors on gastric mucus and epithelial cells. The coccoids with adherence characteristics like the spirals could aid in the pathogenic process of Helicobacter infection. This may probably lead to different clinical outcome of H. pylori associated gastroduodenal disease. PMID:11819557

  5. Spectral characters of lectin saccharide interaction

    NASA Astrophysics Data System (ADS)

    Wang, Deyu; Jiang, Duxiao; Yuan, Chunwei

    1999-09-01

    In this paper we report attempts to directly detect the interaction behavior between erythrocyte and lectin concanavalin a (Con A) as well as phaseolus vulgaris (PHA) on the polystyrene film surface. In the procedure, an optical transducer based reflectance interferometry was set up and used to detect the film thickness change during the lectin adsorption and lectin- erythrocyte interaction. The specific interactions among Con A, PHA and erythrocyte were obtained. The solubility monosaccharide inhibition test confirmed that there is affinity between (alpha) - D-mannose and Con A.

  6. Antiviral lectins as potential HIV microbicides.

    PubMed

    Koharudin, Leonardus M I; Gronenborn, Angela M

    2014-08-01

    A growing class of potential antivirals encompasses carbohydrate-binding proteins, such as antibodies and lectins. They block virus entry into host target cells and halt virus transmission from virus-infected cells to non-infected cells, thereby preventing infection. Here, we review the structural basis for the anti-HIV activity of various lectins, describing their structures and determinants of high-affinity oligosaccharide binding. The mechanism of glycan recognition on the gp120 envelope protein by these antiviral lectins may therefore be exploited for developing agents and alternative strategies to prevent HIV transmission. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Effects of supplementing a basal diet of Chloris gayana hay with one of three protein-rich legume hays of Cassia rotundifolia, Lablab purpureus and Macroptilium atropurpureum forage on some nutritional parameters in goats.

    PubMed

    Mupangwa, J F; Ngongoni, N T; Topps, J H; Hamudikuwanda, H

    2000-08-01

    Growth and digestibility experiments were conducted on growing East African type goats offered Chloris gayana hay supplemented with one of three high-protein (119-128 g CP/kg DM) legume hays, Cassia rotundifolia (cassia), Lablab purpureus (lablab) or Macroptilium atropurpureum (siratro), and crushed maize to investigate the feed intake, digestibility, growth and urinary excretion of purine derivatives. Goats in the supplemented groups had higher total dry matter and nitrogen intakes and higher N retention and body mass gains than unsupplemented counterparts. The digestibility of dry matter, organic matter and neutral detergent fibre were increased by protein supplementation. Animals on supplemented diets had higher fractional outflow rates of particulate matter from the rumen. The production of protein by ruminal microbes and the efficiency of microbial N production were increased by supplementation. It was concluded that a mixture of low-quality grass hay (61.9 CP/kg DM) and either cassia, lablab or siratro hay, and maize grain can provide a productive balanced diet for growing goats.

  8. Legume Lectins: Proteins with Diverse Applications

    PubMed Central

    Lagarda-Diaz, Irlanda; Guzman-Partida, Ana Maria; Vazquez-Moreno, Luz

    2017-01-01

    Lectins are a diverse class of proteins distributed extensively in nature. Among these proteins; legume lectins display a variety of interesting features including antimicrobial; insecticidal and antitumor activities. Because lectins recognize and bind to specific glycoconjugates present on the surface of cells and intracellular structures; they can serve as potential target molecules for developing practical applications in the fields of food; agriculture; health and pharmaceutical research. This review presents the current knowledge of the main structural characteristics of legume lectins and the relationship of structure to the exhibited specificities; provides an overview of their particular antimicrobial; insecticidal and antitumor biological activities and describes possible applications based on the pattern of recognized glyco-targets. PMID:28604616

  9. Galactose-specific seed lectins from Cucurbitaceae.

    PubMed

    Swamy, Musti J; Marapakala, Kavitha; Sultan, Nabil Ali M; Kenoth, Roopa

    2015-01-01

    Lectins, the carbohydrate binding proteins have been studied extensively in view of their ubiquitous nature and wide-ranging applications. As they were originally found in plant seed extracts, much of the work on them was focused on plant seed lectins, especially those from legume seeds whereas much less attention was paid to the lectins from other plant families. During the last two decades many studies have been reported on lectins from the seeds of Cucurbitaceae species. The main focus of the present review is to provide an overview of the current knowledge on these proteins, especially with regard to their physico-chemical characterization, interaction with carbohydrates and hydrophobic ligands, 3-dimensional structure and similarity to type-II ribosome inactivating proteins. The future outlook of research on these galactose-specific proteins is also briefly considered.

  10. Identification of mycobacterial lectins from genomic data.

    PubMed

    Abhinav, K V; Sharma, Alok; Vijayan, M

    2013-04-01

    Sixty-four sequences containing lectin domains with homologs of known three-dimensional structure were identified through a search of mycobacterial genomes. They appear to belong to the β-prism II, the C-type, the Microcystis virdis (MV), and the β-trefoil lectin folds. The first three always occur in conjunction with the LysM, the PI-PLC, and the β-grasp domains, respectively while mycobacterial β-trefoil lectins are unaccompanied by any other domain. Thirty heparin binding hemagglutinins (HBHA), already annotated, have also been included in the study although they have no homologs of known three-dimensional structure. The biological role of HBHA has been well characterized. A comparison between the sequences of the lectin from pathogenic and nonpathogenic mycobacteria provides insights into the carbohydrate binding region of the molecule, but the structure of the molecule is yet to be determined. A reasonable picture of the structural features of other mycobacterial proteins containing one or the other of the four lectin domains can be gleaned through the examination of homologs proteins, although the structure of none of them is available. Their biological role is also yet to be elucidated. The work presented here is among the first steps towards exploring the almost unexplored area of the structural biology of mycobacterial lectins. Copyright © 2012 Wiley Periodicals, Inc.

  11. Animal lectins: potential antitumor therapeutic targets in apoptosis.

    PubMed

    Liu, Zhe; Zhang, Qian; Peng, Hao; Zhang, Wen-zhi

    2012-10-01

    Lectins, a group of carbohydrate-binding proteins ubiquitously distributed into plants and animals, are well-known to have astonishing numerous links to human cancers. In this review, we present a brief outline of the representative animal lectins such as galectins, C-type lectins, and annexins by targeting programmed cell death (or apoptosis) pathways, and also summarize these representative lectins as possible anti-cancer drug targets. Taken together, these inspiring findings would provide a comprehensive perspective for further elucidating the multifaceted roles of animal lectins in apoptosis pathways of cancer, which, in turn, may ultimately help us to exploit lectins for their therapeutic purposes in future drug discovery.

  12. Lectin binding to formalin-fixed paraffin sections.

    PubMed Central

    Leathem, A; Atkins, N

    1983-01-01

    Lectins are potentially useful tools in histopathology for the identification of carbohydrates and distinguishing cells according to their type, differentiation or function. Conjugated to fluorescent or enzyme labels, lectins are simple to use on fresh tissue but fixation and processing sequesters glycoconjugates and dissolves out fat-linked sugars. We describe here the use of labelled antibodies to lectins to localise sites of lectin binding and increase sensitivity, combined with trypsin and neuraminidase to reveal sequestered carbohydrates. Absorbing lectins with appropriate sugars establishes the specificity of binding and allows lectins to be used as sensitive and specific reagents. PMID:6190843

  13. Changes in nonnutritional factors and antioxidant activity during germination of nonconventional legumes.

    PubMed

    Aguilera, Yolanda; Díaz, María Felicia; Jiménez, Tania; Benítez, Vanesa; Herrera, Teresa; Cuadrado, Carmen; Martín-Pedrosa, Mercedes; Martín-Cabrejas, María A

    2013-08-28

    The present study describes the effects of germination on nonnutritional factors and antioxidant activity in the nonconventional legumes Vigna unguiculata (cowpea), Canavalia ensiformis (jack bean), Lablab purpureus (dolichos), and Stizolobium niveum (mucuna). Protease inhibitors and lectins were detected in raw legumes and were significantly decreased during the germination. Regarding total and individual inositol phosphates (IP5-IP3), important reductions of IP6 and high increases in the rest of inositol phosphates were also detected during this process. In addition, total phenols, catechins, and proanthocyanidins increased, accompanied by an overall rise of antioxidant activity (79.6 μmol of Trolox/g of DW in the case of mucuna). Germination has been shown to be a very effective process to reduce nonnutritional factors and increase bioactive phenolic compounds and antioxidant activities of these nonconventional legumes. For this reason, they could be used as ingredients to obtain high-value legume flours for food formulation.

  14. Screening for lectins from basidiomycetes and isolation of Punctularia atropurpurascens lectin.

    PubMed

    Alborés, Silvana; Mora, Paola; Cerdeiras, María Pía; Fraguas, Laura Franco

    2014-02-01

    Aqueous extracts of basidiomycete fungi were screened for the presence of lectins by hemagglutination (HA) assays with mouse, rabbit, and sheep red blood cells. From mycelia and/or fruiting bodies, 23 extracts were prepared; 15 extracts exhibited HA activity towards mouse erythrocytes, with specific activities ranging from 12 to 440 lectin units (LU) mg(-1) protein. In HA inhibition assays, 43 carbohydrates including mono-, di-, tri-, tetrasaccharides, glycoproteins, and polysaccharides were tested as haptens, to determine the saccharide-binding specificities of the lectins. A novel lectin with specificity towards N-acetyl-glucosamine was purified from mycelia of Punctularia atropurpurascens using affinity chromatography on chitosan-Sepharose. The lectin has a subunit molecular mass of 67 kDa determined by SDS-PAGE and a pI of 5.0. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Characterization and antimicrobial activity of lectins from Penicillium sp.

    PubMed

    Singh, R S; Jain, P; Kaur, H P

    2013-11-01

    Ten Penicillium sp. were screened for lectin activity for occurrence of lectins. Mycelial extracts from submerged cultures of P. corylophilum, P. expansum and P. purpurogenum showed agglutination against human (A, B, AB and O), goat, sheep, pig and rabbit erythrocytes. Neuraminidase treatment to human blood- type O erythrocytes substantially increased their agglutinability by all the lectins as compared to untreated erythrocytes. Modification of erythrocyte surfaces by protease increased the lectin titre only of P. corylophilum with no effect on other two lectins. P. corylophilum and P. expansum displayed relatively lower titres in mycelial extracts prepared from agar plate cultures as compared to broth cultures. A panel of sugars was tested for inhibition of lectin activity. All the lectins were found to be specific for asialofetuin, bovine submaxillary mucin, porcine stomach mucin, chondroitin-6-sulphate, D-sucrose and D-glucose. P. corylophilum lectin was expressed (Titre 8) by 5 day old cultures, reaching its maximum level (Titre 32) upon 8 days of cultivation, thereafter declin in lectin activity was observed. P. purpurogenum lectin was expressed by 7-10 days old cultures, while in P. expansum maximum lectin activity was elaborated by 5-8 days old cultures. Lectin extracts from all the three species were found to possess antimicrobial activities. Lectin extracts from the three Penicillium species displayed antifungal activity and antibacterial activity against Gram-negative and Gram-positive bacterial strains.

  16. Epidemiological characterization of Neisseria gonorrhoeae by lectins.

    PubMed Central

    Schalla, W O; Whittington, W L; Rice, R J; Larsen, S A

    1985-01-01

    A total of 101 isolates of penicillinase-producing and non-penicillinase-producing Neisseria gonorrhoeae with known nutritional requirements, plasmid content, and serovars, were examined for lectin agglutination patterns. These isolates were from outbreaks in Georgia, California, Hawaii, and Pennsylvania. Cell suspensions made from 16- to 18-h cultures were mixed with 14 different lectins, and the resultant agglutination patterns were classified as agglutination groups. Among the 101 isolates tested, 24 different agglutination groups were demonstrated. Of the organisms tested, 55% were located in 3 of the 24 groups, and 86% of the isolates reacted with the lectins Trichosanthes kinlowii, Griffonia simplicifolia I, peanut agglutinin, soybean agglutinin, potato agglutinin, and wheat germ agglutinin. One isolate did not react with peanut or potato agglutinin, five isolates lacked reactivity with potato agglutinin, and six isolates did not react with wheat germ agglutinin. Of the wheat germ-negative isolates, four were from Pennsylvania and were identical with regard to auxotype, plasmid content, serovar, and lectin group. The other two wheat germ-negative isolates were from California and were unrelated by the same criteria to the four Pennsylvania isolates and to each other. Among the isolates tested, there were no differences in lectin groups with regard to the sex of the patient. In the Georgia collection, agglutination with one lectin group was confined to isolates of serogroup IA. This association was not observed for the other geographic areas. Some isolates showing identical auxotype, plasmid content, and serovars could be differentiated based on lectin agglutination patterns, whereas other isolates were identical by all testing criteria. PMID:3930560

  17. Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential.

    PubMed

    Gardères, Johan; Bourguet-Kondracki, Marie-Lise; Hamer, Bojan; Batel, Renato; Schröder, Heinz C; Müller, Werner E G

    2015-08-07

    An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial systems. The characterization of the lectins was performed by Edman degradation or mass spectrometry. Regarding their physiological roles, sponge lectins showed to be involved in morphogenesis and cell interaction, biomineralization and spiculogenesis, as well as host defense mechanisms and potentially in the association between the sponge and its microorganisms. In addition, these lectins exhibited a broad range of bioactivities, including modulation of inflammatory response, antimicrobial and cytotoxic activities, as well as anticancer and neuromodulatory activity. In view of their potential pharmacological applications, sponge lectins constitute promising molecules of biotechnological interest.

  18. A rainbow trout lectin with multimeric structure.

    PubMed

    Jensen, L E; Thiel, S; Petersen, T E; Jensenius, J C

    1997-04-01

    A novel lectin has been identified in rainbow trout serum and plasma. The lectin binds to Sepharose (an agarose polymer) in a calcium-dependent manner. Glucose, N-acetyl-glucosamine, mannose, N-acetyl-mannosamine, L-fucose, maltose and alpha-methyl-mannoside are good inhibitors of this binding, whereas glucosamine and D-fucose inhibits to a lesser degree and mannosamine and galactose do not inhibit the binding to Sepharose. When analysed by SDS-PAGE under non-reducing conditions, the lectin appears as a characteristic ladder of bands with approximately 16 kDa between consecutive bands. Upon reduction, the lectin appears as a 16-kDa band. On size-exclusion chromatography of trout serum and plasma, the protein emerges over a broad range corresponding to sizes from about 2000 kDa to less than 200 kDa. The NH2-terminal sequence (AAENRNQXPPG) shows no significant homology with known proteins. Because of the characteristic appearance in non-reducing SDS-PAGE and the lectin activity, we propose to name the protein "ladderlectin."

  19. Antimicrobial and antiparasitic activity of lectins.

    PubMed

    Iordache, Florin; Ionita, Mariana; Mitrea, Liviu Ioan; Fafaneata, Cornelia; Pop, Aneta

    2015-01-01

    Antibiotic resistance is a major problem in current contemporary medicine and it has become a major concern of the 21st century. New resistance mechanisms developed by microorganisms spread greatly, threatening the ability to treat numerous infectious diseases, and increasing the number of nosocomial infections. Besides the role in immunology and glycobiology where they are used as hemaglutinine and identification of complex carbohydrates and glycoconjugates, lectins proved to mediate diversified biological functions like cytotoxicity, complement activation, cell-to-cell and host-pathogen communications, innate immune response, and cell-to-cell signalling. Recently, great interest has been developed for the research and applications of lectins in agriculture and medicine due to their antiparasitic and antimicrobial potentials. This review focuses on the recent data regarding the antimicrobial and antiparasitic activities of lectins, by presenting the role of lectins in host-pathogen interaction and also the cytotoxic effects on microorganisms and parasites. Identification and characterisation of new lectins with antimicrobial activity could serve as a natural alternative for the treatment of infections caused by antibiotic-resistant microorganisms and parasites.

  20. Isolation, characterization and molecular cloning of a leaf-specific lectin from ramsons (Allium ursinum L.).

    PubMed

    Smeets, K; Van Damme, E J; Van Leuven, F; Peumans, W J

    1997-11-01

    Lectins were isolated from roots and leaves of ramsons and compared to the previously described bulb lectins. Biochemical analyses indicated that the root lectins AUAIr and AUAIIr are identical to the bulb lectins AUAI and AUAII, whereas the leaf lectin AUAL has no counterpart in the bulbs. cDNA cloning confirmed that the leaf lectin differs from the bulb lectins. Northern blot analysis further indicated that the leaf lectin is tissue-specifically expressed. Sequence comparisons revealed that the ramsons leaf lectin differs considerably from the leaf lectins of garlic, leek, onion and shallot.

  1. 21 CFR 864.9550 - Lectins and protectins.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Lectins and protectins. 864.9550 Section 864.9550 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... and Blood Products § 864.9550 Lectins and protectins. (a) Identification. Lectins and protectins are...

  2. 21 CFR 864.9550 - Lectins and protectins.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Lectins and protectins. 864.9550 Section 864.9550 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... and Blood Products § 864.9550 Lectins and protectins. (a) Identification. Lectins and protectins are...

  3. A Human Lectin Microarray for Sperm Surface Glycosylation Analysis.

    PubMed

    Sun, Yangyang; Cheng, Li; Gu, Yihua; Xin, Aijie; Wu, Bin; Zhou, Shumin; Guo, Shujuan; Liu, Yin; Diao, Hua; Shi, Huijuan; Wang, Guangyu; Tao, Sheng-Ce

    2016-09-01

    Glycosylation is one of the most abundant and functionally important protein post-translational modifications. As such, technology for efficient glycosylation analysis is in high demand. Lectin microarrays are a powerful tool for such investigations and have been successfully applied for a variety of glycobiological studies. However, most of the current lectin microarrays are primarily constructed from plant lectins, which are not well suited for studies of human glycosylation because of the extreme complexity of human glycans. Herein, we constructed a human lectin microarray with 60 human lectin and lectin-like proteins. All of the lectins and lectin-like proteins were purified from yeast, and most showed binding to human glycans. To demonstrate the applicability of the human lectin microarray, human sperm were probed on the microarray and strong bindings were observed for several lectins, including galectin-1, 7, 8, GalNAc-T6, and ERGIC-53 (LMAN1). These bindings were validated by flow cytometry and fluorescence immunostaining. Further, mass spectrometry analysis showed that galectin-1 binds several membrane-associated proteins including heat shock protein 90. Finally, functional assays showed that binding of galectin-8 could significantly enhance the acrosome reaction within human sperms. To our knowledge, this is the first construction of a human lectin microarray, and we anticipate it will find wide use for a range of human or mammalian studies, alone or in combination with plant lectin microarrays. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. A Human Lectin Microarray for Sperm Surface Glycosylation Analysis *

    PubMed Central

    Sun, Yangyang; Cheng, Li; Gu, Yihua; Xin, Aijie; Wu, Bin; Zhou, Shumin; Guo, Shujuan; Liu, Yin; Diao, Hua; Shi, Huijuan; Wang, Guangyu; Tao, Sheng-ce

    2016-01-01

    Glycosylation is one of the most abundant and functionally important protein post-translational modifications. As such, technology for efficient glycosylation analysis is in high demand. Lectin microarrays are a powerful tool for such investigations and have been successfully applied for a variety of glycobiological studies. However, most of the current lectin microarrays are primarily constructed from plant lectins, which are not well suited for studies of human glycosylation because of the extreme complexity of human glycans. Herein, we constructed a human lectin microarray with 60 human lectin and lectin-like proteins. All of the lectins and lectin-like proteins were purified from yeast, and most showed binding to human glycans. To demonstrate the applicability of the human lectin microarray, human sperm were probed on the microarray and strong bindings were observed for several lectins, including galectin-1, 7, 8, GalNAc-T6, and ERGIC-53 (LMAN1). These bindings were validated by flow cytometry and fluorescence immunostaining. Further, mass spectrometry analysis showed that galectin-1 binds several membrane-associated proteins including heat shock protein 90. Finally, functional assays showed that binding of galectin-8 could significantly enhance the acrosome reaction within human sperms. To our knowledge, this is the first construction of a human lectin microarray, and we anticipate it will find wide use for a range of human or mammalian studies, alone or in combination with plant lectin microarrays. PMID:27364157

  5. Orientation of GST-tagged lectins via in situ surface modification to create an expanded lectin microarray for glycomic analysis.

    PubMed

    Propheter, Daniel C; Mahal, Lara K

    2011-07-01

    Herein we describe the orientation of GST-tagged lectins on NHS-activated slides via a one-step deposition of the protein and a glutathione (GSH) scaffold. This technology overcomes the need for a premade GSH-surface to orient GST-tagged proteins, enabling us to rapidly expand the analytical capacity of lectin microarrays through addition of oriented lectins, while maintaining lectin diversity.

  6. Unfolding energetics and stability of banana lectin.

    PubMed

    Gupta, Garima; Sinha, Sharmistha; Surolia, Avadhesha

    2008-08-01

    The unfolding pathway of banana lectin from Musa paradisiaca was determined by isothermal denaturation induced by the chaotrope GdnCl. The unfolding was found to be a reversible process. The data obtained by isothermal denaturation provided information on conformational stability of banana lectin. The high values of DeltaG of unfolding at various temperatures indicated the strength of intersubunit interactions. It was found that banana lectin is a very stable and denatures at high chaotrope concentrations only. The basis of the stability may be attributed to strong hydrogen bonds of the order 2.5-3.1 A at the dimeric interface along with the presence of water bridges. This is perhaps very unique example in proteins where subunit association is not a consequence of the predominance of hydrophobic interactions.

  7. Diversity of lectins in Macrobrachium rosenbergii and their expression patterns under spiroplasma MR-1008 stimulation.

    PubMed

    Zhu, Huanxi; Du, Jie; Hui, Kai-Min; Liu, Peng; Chen, Jing; Xiu, Yunji; Yao, Wei; Wu, Ting; Meng, Qingguo; Gu, Wei; Ren, Qian; Wang, Wen

    2013-08-01

    Lectins play important roles in crustacean innate immunity through recognition of foreign pathogens. In this study, 20 lectins including C-type lectins [dual-carbohydrate recognition domain (CRD) type and single-CRD type], L-type lectin, and lectin with low-density lipoprotein class A (LDLa) domain were identified from the freshwater prawn Macrobrachium rosenbergii. The tissue distribution and expression patterns of these lectins under spiroplasma strain MR-1008 challenge were investigated. Most of the lectins were found to be mainly distributed in the hepatopancreas. Lectin5, Lectin14, Lectin17, and Lectin18 exhibited the highest expression level in the hemocytes, nerve, intestine, and heart, respectively. MrLec1 to MrLec6 (dual-CRD lectins) in the hepatopancreas were up-regulated by spiroplasma challenge. Single-CRD lectins reached the highest level at 72 h after spiroplasma challenge. Lectin9 and Lectin15 both belong to L-type lectins. At post-spiroplasma challenge, Lectin9 expression was up-regulated, whereas Lectin15 expression was down-regulated. Lectin11 with LDLa domain showed the highest level after 12 h Lectin18 and Lectin20, namely, CD209, were also up-regulated by spiroplasma challenge. Lectin14, a C-type lectin, quickly reached the highest level after 2 h Lectin16 showed the highest level after 72 h Lectin5 reached the highest level in cultured hemocytes after 6 h Lectin17 in the intestine and Lectin14 in the nerve were slightly up-regulated after 6 and 2 h, respectively. Our research results indicate that lectins may play important roles in early or late immune responses against spiroplasma challenge. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Role of Lectins in Plant-Microorganism Interactions

    PubMed Central

    Pueppke, Steven G.; Bauer, Wolfgang D.; Keegstra, Kenneth; Ferguson, Ardene L.

    1978-01-01

    Three different assay procedures have been used to quantitate the levels of soybean (Glycine max [L.] Merr.) lectin in various tissues of soybean plants. The assays used were a standard hemagglutination assay, a radioimmunoassay, and an isotope dilution assay. Most of the lectin in seeds was found in the cotyledons, but lectin was also detected in the embryo axis and the seed coat. Soybean lectin was present in all of the tissues of young seedlings, but decreased as the plants matured and was not detectable in plants older than 2 to 3 weeks. Soybean lectin isolated from seeds of several soybean varieties were identical when compared by several methods. PMID:16660384

  9. Capillary-based lectin affinity electrophoresis for interaction analysis between lectins and glycans.

    PubMed

    Kinoshita, Mitsuhiro; Kakehi, Kazuaki

    2014-01-01

    Capillary affinity electrophoresis (CAE) is a powerful technique for glycan analysis, and one of the analytical approaches for analyzing the interaction between lectins and glycans. The method is based on the high-resolution separation of fluorescently labeled glycans by capillary electrophoresis (CE) with laser-induced fluorescence detection (LIF) in the presence of lectins (or glycan binding proteins). CAE allows simultaneous determination of glycan structures in a complex mixture of glycans. In addition, we can calculate the binding kinetics on a specific glycan in the complex mixture of glycans with a lectin. Here, we show detailed procedures for capillary affinity electrophoresis of fluorescently labeled glycans with lectins using CE-LIF apparatus. Its application to screening a sialic acid binding protein in plant barks is also shown.

  10. Fruit-specific lectins from banana and plantain.

    PubMed

    Peumans, W J; Zhang, W; Barre, A; Houlès Astoul, C; Balint-Kurti, P J; Rovira, P; Rougé, P; May, G D; Van Leuven, F; Truffa-Bachi, P; Van Damme, E J

    2000-09-01

    One of the predominant proteins in the pulp of ripe bananas (Musa acuminata L.) and plantains (Musa spp.) has been identified as a lectin. The banana and plantain agglutinins (called BanLec and PlanLec, respectively) were purified in reasonable quantities using a novel isolation procedure, which prevented adsorption of the lectins onto insoluble endogenous polysaccharides. Both BanLec and PlanLec are dimeric proteins composed of two identical subunits of 15 kDa. They readily agglutinate rabbit erythrocytes and exhibit specificity towards mannose. Molecular cloning revealed that BanLec has sequence similarity to previously described lectins of the family of jacalin-related lectins, and according to molecular modelling studies has the same overall fold and three-dimensional structure. The identification of BanLec and PlanLec demonstrates the occurrence of jacalin-related lectins in monocot species, suggesting that these lectins are more widespread among higher plants than is actually believed. The banana and plantain lectins are also the first documented examples of jacalin-related lectins, which are abundantly present in the pulp of mature fruits but are apparently absent from other tissues. However, after treatment of intact plants with methyl jasmonate, BanLec is also clearly induced in leaves. The banana lectin is a powerful murine T-cell mitogen. The relevance of the mitogenicity of the banana lectin is discussed in terms of both the physiological role of the lectin and the impact on food safety.

  11. Mushroom lectins: specificity, structure and bioactivity relevant to human disease.

    PubMed

    Hassan, Mohamed Ali Abol; Rouf, Razina; Tiralongo, Evelin; May, Tom W; Tiralongo, Joe

    2015-04-08

    Lectins are non-immunoglobulin proteins that bind diverse sugar structures with a high degree of selectivity. Lectins play crucial role in various biological processes such as cellular signaling, scavenging of glycoproteins from the circulatory system, cell-cell interactions in the immune system, differentiation and protein targeting to cellular compartments, as well as in host defence mechanisms, inflammation, and cancer. Among all the sources of lectins, plants have been most extensively studied. However, more recently fungal lectins have attracted considerable attention due to their antitumor, antiproliferative and immunomodulatory activities. Given that only 10% of mushroom species are known and have been taxonomically classified, mushrooms represent an enormous unexplored source of potentially useful and novel lectins. In this review we provide an up-to-date summary on the biochemical, molecular and structural properties of mushroom lectins, as well as their versatile applications specifically focusing on mushroom lectin bioactivity.

  12. Comprehensive list of lectins: origins, natures, and carbohydrate specificities.

    PubMed

    Kobayashi, Yuka; Tateno, Hiroaki; Ogawa, Haruko; Yamamoto, Kazuo; Hirabayashi, Jun

    2014-01-01

    More than 100 years have passed since the first lectin ricin was discovered. Since then, a wide variety of lectins (lect means "select" in Latin) have been isolated from plants, animals, fungi, bacteria, as well as viruses, and their structures and properties have been characterized. At present, as many as 48 protein scaffolds have been identified as functional lectins from the viewpoint of three-dimensional structures as described in this chapter. In this chapter, representative 53 lectins are selected, and their major properties that include hemagglutinating activity, mitogen activity, blood group specificity, molecular weight, metal requirement, and sugar specificities are summarized as a comprehensive table. The list will provide a practically useful, comprehensive list for not only experienced lectin users but also many other non-expert researchers, who are not familiar to lectins and, therefore, have no access to advanced lectin biotechnologies described in other chapters.

  13. Diversity and multiple functions of lectins in shrimp immunity.

    PubMed

    Wang, Xian-Wei; Wang, Jin-Xing

    2013-01-01

    Lectins play important roles in many biological processes, including protein trafficking, cell signaling, pathogen recognition, as effector molecules, and so on, because of their capacity to bind carbohydrates. Presently, seven groups of lectins have been identified in shrimp: C-type, L-type, P-type, M-type, fibrinogen-like domain lectins, galectins, and calnexin/calreticulin. These lectins have different structures, diverse expression patterns, and multiple functions in the shrimp immune response. This review summarizes the research progress and analyzes the diversity of shrimp lectins, focusing mainly on the C-type lectin family. Shrimp C-type lectins show considerable diversity in their domain architectures, sugar substrates, tissue distributions, expression patterns responding to pathogen challenge and functions in shrimp immunity. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Mushroom Lectins: Specificity, Structure and Bioactivity Relevant to Human Disease

    PubMed Central

    Hassan, Mohamed Ali Abol; Rouf, Razina; Tiralongo, Evelin; May, Tom W.; Tiralongo, Joe

    2015-01-01

    Lectins are non-immunoglobulin proteins that bind diverse sugar structures with a high degree of selectivity. Lectins play crucial role in various biological processes such as cellular signaling, scavenging of glycoproteins from the circulatory system, cell–cell interactions in the immune system, differentiation and protein targeting to cellular compartments, as well as in host defence mechanisms, inflammation, and cancer. Among all the sources of lectins, plants have been most extensively studied. However, more recently fungal lectins have attracted considerable attention due to their antitumor, antiproliferative and immunomodulatory activities. Given that only 10% of mushroom species are known and have been taxonomically classified, mushrooms represent an enormous unexplored source of potentially useful and novel lectins. In this review we provide an up-to-date summary on the biochemical, molecular and structural properties of mushroom lectins, as well as their versatile applications specifically focusing on mushroom lectin bioactivity. PMID:25856678

  15. Efficacy of glycoprotein enrichment by microscale lectin affinity chromatography.

    PubMed

    Madera, Milan; Mann, Benjamin; Mechref, Yehia; Novotny, Milos V

    2008-08-01

    Reproducible and efficient affinity enrichment is increasingly viewed as an essential step in many investigations leading to the discovery of new biomarkers. In this work, we have evaluated the repeatability of lectin enrichment of glycoproteins from human blood serum through both qualitative and quantitative proteomic approaches. In a comprehensive evaluation of lectin binding, we have performed 30 separate microscale lectin affinity chromatography experiments, followed by a conventional sample purification, and LC-MS/MS analysis of the enriched glycoproteins. Two lectin affinity matrixes, both with Con A lectin, immobilized to the same solid support but differing in the amount of immobilized lectin, were investigated to characterize their binding properties. Both qualitative and quantitative data indicate acceptable repeatability and binding efficiency for the lectin materials received from two different commercial sources.

  16. Lectins from tropical sponges. Purification and characterization of lectins from genus Aplysina.

    PubMed

    Miarons, P B; Fresno, M

    2000-09-22

    Only a few animal phyla have been screened for the presence and distribution of lectins. Probably the most intensively studied group is the mollusk. In this investigation, 22 species from 12 families of tropical sponges collected in Los Roques National Park (Venezuela) were screened for the presence of lectins. Nine saline extracts exhibited strong hemagglutinating activity against pronase-treated hamster red blood cells; five of these reacted against rabbit red blood cells, four with trypsin-treated bovine red blood cells, and five with human red blood cells regardless of the blood group type. Extracts from the three species studied from genus Aplysina (archeri, lawnosa, and cauliformis) were highly reactive and panagglutinating against the panel of red blood cells tested. The lectins from A. archeri and A. lawnosa were purified to homogeneity by ammonium sulfate fractionation, affinity chromatography on p-aminobenzyl-beta-1-thiogalactopyranoside-agarose, and gel filtration chromatography. Both lectins exhibited a native molecular mass of 63 kDa and by SDS-polyacrylamide gel electrophoresis under reducing conditions have an apparent molecular mass of 16 kDa, thus suggesting they occur as homotetramers. The purified lectins contain 3-4 mol of divalent cation per molecule, which are essential for their biological activity. Hapten inhibition of hemagglutination was carried out to define the sugar binding specificity of the purified A. archeri lectin. The results indicate a preference of the lectin for nonreducing beta-linked d-Gal residues being the best inhibitors of red blood cells binding methyl-beta-d-Gal and thiodigalactoside (Gal beta 1-4-thiogalactopyranoside). The behavior of several glycans on immobilized lectin affinity chromatography confirmed and extended the specificity data obtained by hapten inhibition.

  17. Lectin-mediated drug delivery: influence of mucin on cytoadhesion of plant lectins in vitro.

    PubMed

    Wirth, M; Gerhardt, K; Wurm, C; Gabor, F

    2002-02-19

    As the mucous layer represents the first barrier to peroral lectin-mediated drug delivery, the influence of mucin on the cytoadhesive properties of lectins was studied in vitro by establishing a rapid and simple microplate format assay using pig gastric mucin (PGM) for coating the wells. The lectin-binding capacity of mucin followed the order WGA>UEA-I>LCA=STL>PNA>DBA. The PGM-binding of wheat germ agglutinin (WGA) was strongly dependent on pH being highest at pH 5.0. In comparison, PGM-binding of WGA was about 15% at gastric pH and 60-70% at intestinal pH. This points to unimpeded gastric transit of WGA-grafted formulations and favorable conditions within the intestine for binding to mucus coated enterocytes. Moreover the WGA-PGM interaction was concentration-dependent, specific and fully reversible. According to a competitive assay in the presence of Caco-2 monolayers, the PGM-binding of WGA was saturated and influenced by the lectin-concentration yielding 28% Caco-2 bound WGA (125 ng WGA/0.29 cm(2) monolayer) and 68% Caco-2 bound WGA (4 microg WGA/0.29 cm(2) monolayer), respectively. Following on from these results, lectins are expected to suffer at least partially from premature inactivation by shed off mucus like bioadhesives of the first generation, however initial but reversible mucus-binding of lectins offers partititioning to the cell membrane followed by uptake into the enterocyte.

  18. Structural analysis of a Dioclea sclerocarpa lectin: Study on the vasorelaxant properties of Dioclea lectins.

    PubMed

    Barroso-Neto, Ito L; Delatorre, Plinio; Teixeira, Claudener S; Correia, Jorge L A; Cajazeiras, João B; Pereira, Ronniery I; Nascimento, Kyria S; Laranjeira, Eva P P; Pires, Alana F; Assreuy, Ana M S; Rocha, Bruno A M; Cavada, Benildo S

    2016-01-01

    Lectins are proteins that show a variety of biological activities. However, they share in common at least one domain capable of recognizing specific carbohydrates reversibly without changing its structure. The legume lectins family is the most studied family of plant lectins, in particular the Diocleinae subtribe, which possesses high degree of structural similarity, but variable biological activities. This variability lies in small differences that can be analyzed in studies based on structures. In particular, Dioclea sclerocarpa seed lectin (DSL) presents low ability to relax endothelialized rat aorta in comparison with other Dioclea lectins such as Dioclea violacea (DVL), Dioclea virgata (DvirL) and Dioclea rostrata (DRL). The DSL relaxation mechanism relies on nitric oxide production and carbohydrate recognition domain (CRD). This feature can be explained by structural differences, since DSL has a carbohydrate recognition domain design less favorable. In addition, the presence of a glutamate residue at position 205 proved to be a decisive factor for the low relaxant effect of Dioclea lectins. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Development and Applications of the Lectin Microarray.

    PubMed

    Hirabayashi, Jun; Kuno, Atsushi; Tateno, Hiroaki

    2015-01-01

    The lectin microarray is an emerging technology for glycomics. It has already found maximum use in diverse fields of glycobiology by providing simple procedures for differential glycan profiling in a rapid and high-throughput manner. Since its first appearance in the literature in 2005, many application methods have been developed essentially on the same platform, comprising a series of glycan-binding proteins immobilized on an appropriate substrate such as a glass slide. Because the lectin microarray strategy does not require prior liberation of glycans from the core protein in glycoprotein analysis, it should encourage researchers not familiar with glycotechnology to use glycan analysis in future work. This feasibility should provide a broader range of experimental scientists with good opportunities to investigate novel aspects of glycoscience. Applications of the technology include not only basic sciences but also the growing fields of bio-industry. This chapter describes first the essence of glycan profiling and the basic fabrication of the lectin microarray for this purpose. In the latter part the focus is on diverse applications to both structural and functional glycomics, with emphasis on the wide applicability now available with this new technology. Finally, the importance of developing advanced lectin engineering is discussed.

  20. BAD-lectins: boronic acid-decorated lectins with enhanced binding affinity for the selective enrichment of glycoproteins.

    PubMed

    Lu, Ying-Wei; Chien, Chih-Wei; Lin, Po-Chiao; Huang, Li-De; Chen, Chang-Yang; Wu, Sz-Wei; Han, Chia-Li; Khoo, Kay-Hooi; Lin, Chun-Cheng; Chen, Yu-Ju

    2013-09-03

    The weak and variable binding affinities exhibited by lectin-carbohydrate interactions have often compromised the practical utility of lectin in capturing glycoproteins for glycoproteomic applications. We report here the development and applications of a new type of hybrid biomaterial, namely a boronic acid-decorated lectin (BAD-lectin), for efficient bifunctional glycoprotein labeling and enrichment. Our binding studies showed an enhanced affinity by BAD-lectin, likely to be mediated via the formation of boronate ester linkages between the lectin and glycan subsequent to the initial recognition process and thus preserving its glycan-specificity. Moreover, when attached to magnetic nanoparticles (BAD-lectin@MNPs), 2 to 60-fold improvement on detection sensitivity and enrichment efficiency for specific glycoproteins was observed over the independent use of either lectin or BA. Tested at the level of whole cell lysates for glycoproteomic applications, three different types of BAD-lectin@MNPs exhibited excellent specificities with only 6% overlapping among the 295 N-linked glycopeptides identified. As many as 236 N-linked glycopeptides (80%) were uniquely identified by one of the BAD-lectin@MNPs. These results indicated that the enhanced glycan-selective recognition and binding affinity of BAD-lectin@MNPs will facilitate a complementary identification of the under-explored glycoproteome.

  1. Lectins, Interconnecting Proteins with Biotechnological/Pharmacological and Therapeutic Applications

    PubMed Central

    Silva, Priscila Marcelino dos Santos

    2017-01-01

    Lectins are proteins extensively used in biomedical applications with property to recognize carbohydrates through carbohydrate-binding sites, which identify glycans attached to cell surfaces, glycoconjugates, or free sugars, detecting abnormal cells and biomarkers related to diseases. These lectin abilities promoted interesting results in experimental treatments of immunological diseases, wounds, and cancer. Lectins obtained from virus, microorganisms, algae, animals, and plants were reported as modulators and tool markers in vivo and in vitro; these molecules also play a role in the induction of mitosis and immune responses, contributing for resolution of infections and inflammations. Lectins revealed healing effect through induction of reepithelialization and cicatrization of wounds. Some lectins have been efficient agents against virus, fungi, bacteria, and helminths at low concentrations. Lectin-mediated bioadhesion has been an interesting characteristic for development of drug delivery systems. Lectin histochemistry and lectin-based biosensors are useful to detect transformed tissues and biomarkers related to disease occurrence; antitumor lectins reported are promising for cancer therapy. Here, we address lectins from distinct sources with some biological effect and biotechnological potential in the diagnosis and therapeutic of diseases, highlighting many advances in this growing field. PMID:28367220

  2. Sensing lectin-glycan interactions using lectin super-microarrays and glycans labeled with dye-doped silica nanoparticles.

    PubMed

    Wang, Xin; Matei, Elena; Deng, Lingquan; Koharudin, Leonardus; Gronenborn, Angela M; Ramström, Olof; Yan, Mingdi

    2013-09-15

    A new microarray platform, based on lectin super-microarrays and glycans labeled with dye-doped nanoparticles, has been developed to study glycan-lectin interactions. Glycan ligands were conjugated onto fluorescein-doped silica nanoparticles (FSNPs) using a general photocoupling chemistry to afford FSNP-labeled glycan probes. Lectins were printed on epoxy slides in duplicate sets to generate lectin super-microarrays where multiple assays could be carried out simultaneously in each lectin microarray. Thus, the lectin super-microarray was treated with FSNP-labeled glycans to screen for specific binding pairs. Furthermore, a series of ligand competition assays were carried out on a single lectin super-microarray to generate the dose-response curve for each glycan-lectin pair, from which the apparent affinity constants were obtained. Results showed 4-7 orders of magnitude increase in affinity over the free glycans with the corresponding lectins. Thus, the glycan epitope structures having weaker affinity than the parent glycans could be readily identified and analyzed from the lectin super-microarrays. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. A profile of protein-protein interaction: Crystal structure of a lectin-lectin complex.

    PubMed

    Surya, Sukumaran; Abhilash, Joseph; Geethanandan, Krishnan; Sadasivan, Chittalakkottu; Haridas, Madhathilkovilakathu

    2016-06-01

    Proteins may utilize complex networks of interactions to create/proceed signaling pathways of highly adaptive responses such as programmed cell death. Direct binary interactions study of proteins may help propose models for protein-protein interaction. Towards this goal we applied a combination of thermodynamic kinetics and crystal structure analyses to elucidate the complexity and diversity in such interactions. By determining the heat change on the association of two galactose-specific legume lectins from Butea monosperma (BML) and Spatholobus parviflorus (SPL) belonging to Fabaceae family helped to compute the binding equilibrium. It was extended further by X-ray structural analysis of BML-SPL binary complex. In order to chart the proteins interacting mainly through their interfaces, identification of the nature of forces which stabilized the association of the lectin-lectin complex was examined. Comprehensive analysis of the BMLSPL complex by isothermal titration calorimetry and X-ray crystal structure threw new light on the lectin-lectin interactions suggesting of their use in diverse areas of glycobiology.

  4. Using Single Lectins to Enrich Glycoproteins in Conditioned Media.

    PubMed

    Sethi, Manveen K; Fanayan, Susan

    2015-08-03

    Lectins are sugar-binding proteins that can recognize and bind to carbohydrates conjugated to proteins and lipids. Coupled with mass spectrometry technologies, lectin affinity chromatography is becoming a popular approach for identification and quantification of glycoproteins in complex samples such as blood, tumor tissues, and cell lines. Given the commercial availability of a large number of lectins that recognize diverse sugar structures, it is now possible to isolate and study glycoproteins for biological and medical research. This unit provides a general guide to single-lectin-based enrichment of glycoproteins from serum-free conditioned media. Due to the unique carbohydrate specificity of most lectins and the complexity of the samples, optimization steps may be required to evaluate different elution buffers and methods as well as binding conditions, for each lectin, for optimal recovery of bound glycoproteins.

  5. Lectin cDNA and transgenic plants derived therefrom

    DOEpatents

    Raikhel, N.V.

    1994-01-04

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties. GOVERNMENT RIGHTS This application was funded under Department of Energy Contract DE-AC02-76ER01338. The U.S. Government has certain rights under this application and any patent issuing thereon. .

  6. Lectin cDNA and transgenic plants derived therefrom

    DOEpatents

    Raikhel, Natasha V.

    1994-01-04

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties. GOVERNMENT RIGHTS This application was funded under Department of Energy Contract DE-AC02-76ER01338. The U.S. Government has certain rights under this application and any patent issuing thereon.

  7. Soluble Lectins: A New Class of Extracellular Proteins

    NASA Astrophysics Data System (ADS)

    Barondes, Samuel H.

    1984-03-01

    Soluble lectins of cellular slime molds and vertebrates are present at extracellular sites in the developing or adult tissues that make them. Some lectins are concentrated around cell groups, as in extracellular matrix or elastic fibers. Others are at the interface between cells and the external environment, as in much or slime. Specific glycoproteins, proteoglycans, or polysaccharides that bind these endogenous lectins may also be present at these sites. Interactions between the lectins and glycoconjugates appear to play a role in shaping extracellular environments.

  8. Lectins discriminate between pathogenic and nonpathogenic South American trypanosomes

    SciTech Connect

    de Miranda Santos, I.K.; Pereira, M.E.

    1984-09-01

    Cell surface carbohydrates of Trypanosoma cruzi, Trypanosoma rangeli, and Trypanosoma conorhini were analyzed by a micro-agglutination assay employing 27 highly purified lectins and by binding assays using various /sup 125/I-labeled lectins. The following seven lectins discriminated between the trypanosomes: 1) tomato lectin (an N-acetyl-D-glucosamine-binding protein), both in purified form and as crude tomato juice; 2) Bauhinea purpurea and Sophora japonica lectins (both N-acetyl-D-galactosamine-binding proteins), which selectively agglutinated T. cruzi; 3) Vicia villosa (an N-acetyl-D-galactosamine-binding protein) which was specific for T. rangeli; 4) peanut lectin (a D-galactose-binding protein) both in purified form and as crude saline extract; and 5) Ulex europaeus and Lotus tetragonolobus (both L-fucose-binding proteins) lectins which reacted only with T. conorhini. Binding studies with 125I-labeled lectins were performed to find whether unagglutinated cells of the three different species of trypanosomes might have receptors for these lectins, in which case absence of agglutination could be due to a peculiar arrangement of the receptors. These assays essentially confirmed the agglutination experiments.

  9. Lectins discriminate between pathogenic and nonpathogenic South American trypanosomes.

    PubMed

    de Miranda Santos, I K; Pereira, M E

    1984-09-01

    Cell surface carbohydrates of Trypanosoma cruzi, Trypanosoma rangeli, and Trypanosoma conorhini were analyzed by a micro-agglutination assay employing 27 highly purified lectins and by binding assays using various 125I-labeled lectins. The following seven lectins discriminated between the trypanosomes: 1) tomato lectin (an N-acetyl-D-glucosamine-binding protein), both in purified form and as crude tomato juice; 2) Bauhinea purpurea and Sophora japonica lectins (both N-acetyl-D-galactosamine-binding proteins), which selectively agglutinated T. cruzi; 3) Vicia villosa (an N-acetyl-D-galactosamine-binding protein) which was specific for T. rangeli; 4) peanut lectin (a D-galactose-binding protein) both in purified form and as crude saline extract; and 5) Ulex europaeus and Lotus tetragonolobus (both L-fucose-binding proteins) lectins which reacted only with T. conorhini. Binding studies with 125I-labeled lectins were performed to find whether unagglutinated cells of the three different species of trypanosomes might have receptors for these lectins, in which case absence of agglutination could be due to a peculiar arrangement of the receptors. These assays essentially confirmed the agglutination experiments.

  10. A thermostable lectin from the rhizomes of Kaempferia parviflora.

    PubMed

    Konkumnerd, Wichchulada; Karnchanatat, Aphichart; Sangvanich, Polkit

    2010-08-30

    Kaempferia parviflora, or black galingale (Kra-Chai-Dam), belongs to the Zingiberaceae family and is used as both a food ingredient and a medicinal plant. There are diverse reports on the biological activities of compounds extracted from the plant, such as antimalarial, antifungal and an effective sexual-enhancing role, but not on the lectins. A lectin was isolated from the rhizomes of Kaempferia parviflora using affinity chromatography on Concanavalin A followed by gel filtration chromatography on Sephacryl S-100. The molecular weight of the purified lectin was about 41.7 kDa. This lectin showed haemagglutinating activity against erythrocytes from several sources, with the highest level being against those from rabbits. Moreover, the lectin was thermostable, with significant haemagglutinating activity detectable up to 75 degrees C. The results of trypsin digestion and liquid chromatography/tandem mass spectrometry analysis suggested that this protein could be a member of the lectin/endochitnase1 family. A lectin that showed thermotolerant haemagglutinating activity against erythrocytes from several sources was successfully purified from K. paviflora rhizomes. Peptide sequence analysis indicated that this lectin is similar to lectin/endochitinase 1 (Urtica dioica) or Hevein-like protein (Hevea brasiliensis). Copyright (c) 2010 Society of Chemical Industry.

  11. Archeal lectins: An identification through a genomic search.

    PubMed

    Abhinav, K V; Samuel, Ebenezer; Vijayan, M

    2016-01-01

    Forty-six lectin domains which have homologues among well established eukaryotic and bacterial lectins of known three-dimensional structure, have been identified through a search of 165 archeal genomes using a multipronged approach involving domain recognition, sequence search and analysis of binding sites. Twenty-one of them have the 7-bladed β-propeller lectin fold while 16 have the β-trefoil fold and 7 the legume lectin fold. The remainder assumes the C-type lectin, the β-prism I and the tachylectin folds. Acceptable models of almost all of them could be generated using the appropriate lectins of known three-dimensional structure as templates, with binding sites at one or more expected locations. The work represents the first comprehensive bioinformatic study of archeal lectins. The presence of lectins with the same fold in all domains of life indicates their ancient origin well before the divergence of the three branches. Further work is necessary to identify archeal lectins which have no homologues among eukaryotic and bacterial species. © 2015 Wiley Periodicals, Inc.

  12. Functional assay using lectin gene targeting technologies (over-expression).

    PubMed

    Nonaka, Motohiro; Kawasaki, Toshisuke

    2014-01-01

    Function of lectin depends on its amino acid sequence of carbohydrate-recognition domain (CRD), conformation, and extracellular/intracellular localization. Altering lectin gene expression by over-expression or knockdown is a powerful tool for analyzing its cellular function. Here, we describe a method of lectin gene over-expression, taking a C-type lectin, mannan-binding protein (MBP), as an example. Carbohydrate-binding ability of MBP, its subcellular localization, and functional co-localization with ligand glycoprotein are assayed comparing with an inactive mutant MBP.

  13. Role of bean lectins inRhizobium phaseoli-Phaseolus vulgaris interactions. Some properties of lectins from two bean cultivars.

    PubMed

    Mariño, G B; Boiardi, J L

    1992-11-01

    Lectins from twoPhaseolus vulgaris L. cultivars were isolated and purified by salt fractionation, affinity chromatography and gel permeation chromatography. The cultivars used were: 'alubia', with a low-nodulating ability, and Bat 76 with a good symbiotic aptitude. Differences in properties of the two lectins were noted: 'alubia' lectin gave only one peak with haemagglutinating activity following gel permeation chromatography while Bat 76 yielded two active peaks, although both lectins had several bands of about 30 kDa following gel electrophoresis, Bat 76 lecting had three bands of about 50 kDa which were not present in 'alubia' and red kidney bean lectins. Peptide-mapping, by limited proteolysis and two dimensional gel electrophoresis, also showed differences between the lectins which are therefore judged to be different.

  14. Algal Lectins as Potential HIV Microbicide Candidates

    PubMed Central

    Huskens, Dana; Schols, Dominique

    2012-01-01

    The development and use of topical microbicides potentially offers an additional strategy to reduce the spread of the Human Immunodeficiency Virus (HIV). Carbohydrate-binding agents (CBAs) that show specificity for high mannose carbohydrates on the surface of the heavily glycosylated envelope of HIV are endowed with potent anti-HIV activity. In fact, a number of algal lectins such as cyanovirin-N, microvirin, microcystis viridis lectin, scytovirin, Oscillatoria agardhii agglutinin and griffithsin are considered as potential microbicide candidates to prevent the sexual transmission of HIV through topical applications. They not only inhibit infection of cells by cell-free virus but they can also efficiently prevent virus transmission from virus-infected cells to uninfected CD4+ target T-lymphocytes and DC-SIGN-directed capture of HIV-1 and transmission to CD4+ T lymphocytes. This review focuses on the structural properties and carbohydrate specificity of these algal lectins, their antiviral activity against HIV and several other enveloped viruses, their safety profile and viral resistance patterns. PMID:22851920

  15. Plant Lectins and Lectin Receptor-Like Kinases: How Do They Sense the Outside?

    PubMed

    Bellande, Kevin; Bono, Jean-Jacques; Savelli, Bruno; Jamet, Elisabeth; Canut, Hervé

    2017-05-31

    Lectins are fundamental to plant life and have important roles in cell-to-cell communication; development and defence strategies. At the cell surface; lectins are present both as soluble proteins (LecPs) and as chimeric proteins: lectins are then the extracellular domains of receptor-like kinases (LecRLKs) and receptor-like proteins (LecRLPs). In this review; we first describe the domain architectures of proteins harbouring G-type; L-type; LysM and malectin carbohydrate-binding domains. We then focus on the functions of LecPs; LecRLKs and LecRLPs referring to the biological processes they are involved in and to the ligands they recognize. Together; LecPs; LecRLKs and LecRLPs constitute versatile recognition systems at the cell surface contributing to the detection of symbionts and pathogens; and/or involved in monitoring of the cell wall structure and cell growth.

  16. A comparison of tomato (Lycopersicon esculentum) lectin with its deglycosylated derivative.

    PubMed

    Kilpatrick, D C; Graham, C; Urbaniak, S J; Jeffree, C E; Allen, A K

    1984-06-15

    A deglycosylated derivative of tomato (Lycopersicon esculentum) lectin was prepared with the use of trifluoromethanesulphonic acid. Its properties were generally similar to those of the native lectin, but differences were evident in terms of relative agglutinating activity towards sheep, (untreated) human and trypsin-treated human erythrocytes. The two forms of tomato lectin were used in conjunction with a battery of specific antisera to investigate structural relatedness among solanaceous lectins. Immunological cross-reactivity between tomato, potato and Datura lectins depends on the integrity of the glycosylated region of those lectins; that between Datura lectin and other seed lectins, however, has a separate structural basis.

  17. Inhibition of Haemonchus contortus larval development by fungal lectins.

    PubMed

    Heim, Christian; Hertzberg, Hubertus; Butschi, Alex; Bleuler-Martinez, Silvia; Aebi, Markus; Deplazes, Peter; Künzler, Markus; Štefanić, Saša

    2015-08-19

    Lectins are carbohydrate-binding proteins that are involved in fundamental intra- and extracellular biological processes. They occur ubiquitously in nature and are especially abundant in plants and fungi. It has been well established that certain higher fungi produce lectins in their fruiting bodies and/or sclerotia as a part of their natural resistance against free-living fungivorous nematodes and other pests. Despite relatively high diversity of the glycan structures in nature, many of the glycans targeted by fungal lectins are conserved among organisms of the same taxon and sometimes even among different taxa. Such conservation of glycans between free-living and parasitic nematodes is providing us with a useful tool for discovery of novel chemotherapeutic and vaccine targets. In our study, a subset of fungal lectins emanating from toxicity screens on Caenorhabditis elegans was tested for their potential to inhibit larval development of Haemonchus contortus. The effect of Coprinopsis cinerea lectins - CCL2, CGL2, CGL3; Aleuria aurantia lectin - AAL; Marasmius oreades agglutinin - MOA; and Laccaria bicolor lectin - Lb-Tec2, on cultivated Haemonchus contortus larval stages was investigated using a larval development test (LDT). To validate the results of the toxicity assay and determine lectin binding capacity to the nematode digestive tract, biotinylated versions of lectins were fed to pre-infective larval stages of H. contortus and visualized by fluorescent microscopy. Lectin histochemistry on fixed adult worms was performed to investigate the presence and localisation of lectin binding sites in the disease-relevant developmental stage. Using an improved larval development test we found that four of the six tested lectins: AAL, CCL2, MOA and CGL2, exhibited a dose-dependent toxicity in LDT, as measured by the number of larvae developing to the L3 stage. In the case of AAL, CGL2 and MOA lectin, doses as low as 5 μg/ml caused >95 % inhibition of larval

  18. Cell-to-cell binding induced by different lectins.

    PubMed

    Rutishauser, U; Sachs, L

    1975-05-01

    The cell-to-cell binding induced by concanavalin A (Con A) and the lectins from wheatgerm, soybean, and waxbean has been analyzed by measuring the ability of single cells to bind to lectin-coated cells immobilized on nylon fibers. The cells used were lymphoma, myeloid leukemia, and normal fibroblast cells. With all lectins, cell-to-cell binding was inhibited if both cells were prefixed with glutaraldehyde. However, in most cases cell-to-cell binding was enhanced when only the lectin-coated cell was prefixed. With normal fibroblasts, treatment of either one or both cells with trypsin enhanced the cell-to-cell binding induced by Con A and the wheatgerm lectin. Neuraminidase, which increases the number of receptors for soybean agglutinin, increased cell-to-cell binding only if both cells were treated. Although cell-to-cell binding induced by the lectins from soybean and wheatgerm could be partially reversed by the appropriate competitive saccharide inhibitor, binding induced by Con A could not be reversed. The experiments indicate that cell-to-cell binding induced by a lectin can be prevented by an insufficient density of receptors for the lectin, insufficient receptor mobility, or induced clustering of receptors. These effects can explain the differences in cell-to-cell binding and agglutination observed with different cell types and lectins. They also suggest that cell-to-cell binding induced by different lectins with a variety of cell types is initiated by a mechanism involving the alignment of complementary receptors on the colliding cells for the formation of multiple cell-to-lectin-to-cell bridges.

  19. Characterization of mannose binding lectin from channel catfish Ictalurus punctatus

    USDA-ARS?s Scientific Manuscript database

    Mannose-binding lectin (MBL) is an important component of innate immunity capable of activating the lectin pathway of the complement system. A MBL gene was isolated from channel catfish (Ictalurus punctatus). The deduced protein contains a canonical collagen-like domain, a carbohydrate recognition d...

  20. Modulation of glycan detection on specific glycoproteins by lectin multimerization.

    PubMed

    Cao, Zheng; Partyka, Katie; McDonald, Mitchell; Brouhard, Elizabeth; Hincapie, Marina; Brand, Randall E; Hancock, William S; Haab, Brian B

    2013-02-05

    Improved methods for studying glycans could spur significant advances in the understanding and application of glycobiology. The use of affinity reagents such as lectins and glycan-binding antibodies is a valuable complement to methods involving mass spectrometry and chromatography. Many lectins, however, are not useful as analytic tools due to low affinity in vitro. As an approach to increasing lectin avidity to targeted glycans, we tested the use of lectin multimerization. Several biotinylated lectins were linked together through streptavidin interactions. The binding of certain lectins for purified glycoproteins and glycoproteins captured directly out of biological solutions was increased using multimerization, resulting in the detection of lower concentrations of glycoprotein than possible using monomeric detection. The analysis of glycoproteins in plasma samples showed that the level of binding enhancement through multimerization was not equivalent across patient samples. Wheat germ agglutinin (WGA) reactive glycans on fibronectin and thrombospondin-5 were preferentially bound by multimers in pancreatic cancer patient samples relative to control samples, suggesting a cancer-associated change in glycan density that could be detected only through lectin multimerization. This strategy could lead to the more sensitive and informative detection of glycans in biological samples and a broader spectrum of lectins that are useful as analytical reagents.

  1. Plant Lectins: Wheat Defense Strategy Against Hessian Fly

    USDA-ARS?s Scientific Manuscript database

    Plants produce a variety of defense proteins, including lectins in response to attack by phytophagous insects. Ultrastructural studies reveal that binding to insect gut structures and resistance to proteolytic degradation by insect digestive enzymes are the two main prerequisites for the lectins to...

  2. Isothermal calorimetric analysis of lectin-sugar interaction.

    PubMed

    Takeda, Yoichi; Matsuo, Ichiro

    2014-01-01

    Isothermal titration calorimetry (ITC) is a powerful tool for analyzing lectin-glycan interactions because it can measure the binding affinity and thermodynamic properties such as ∆H and ΔS in a single experiment without any chemical modification or immobilization. Here we describe a method for preparing glycan and lectin solution to minimize the buffer mismatch, setting parameters, and performing experiments.

  3. Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential

    PubMed Central

    Gardères, Johan; Bourguet-Kondracki, Marie-Lise; Hamer, Bojan; Batel, Renato; Schröder, Heinz C.; Müller, Werner E. G.

    2015-01-01

    An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial systems. The characterization of the lectins was performed by Edman degradation or mass spectrometry. Regarding their physiological roles, sponge lectins showed to be involved in morphogenesis and cell interaction, biomineralization and spiculogenesis, as well as host defense mechanisms and potentially in the association between the sponge and its microorganisms. In addition, these lectins exhibited a broad range of bioactivities, including modulation of inflammatory response, antimicrobial and cytotoxic activities, as well as anticancer and neuromodulatory activity. In view of their potential pharmacological applications, sponge lectins constitute promising molecules of biotechnological interest. PMID:26262628

  4. Lectin-binding properties of Aeromonas caviae strains

    PubMed Central

    Rocha-de-Souza, Cláudio M.; Hirata-Jr, Raphael; Mattos-Guaraldi, Ana L.; Freitas-Almeida, Angela C.; Andrade, Arnaldo F. B.

    2008-01-01

    The cell surface carbohydrates of four strains of Aeromonas caviae were analyzed by agglutination and lectin-binding assays employing twenty highly purified lectins encompassing all sugar specificities. With the exception of L-fucose and sialic acid, the sugar residues were detected in A. caviae strains. A marked difference, however, in the pattern of cell surface carbohydrates in different A. caviae isolates was observed. Specific receptors for Tritricum vulgaris (WGA), Lycopersicon esculentum (LEL) and Solanum tuberosum (STA) (D-GlcNAc-binding lectins) were found only in ATCC 15468 strain, whereas Euonymus europaeus (EEL, D-Gal-binding lectin) sites were present exclusively in AeQ32 strain, those for Helix pomatia (HPA, D-GalNAc-binding lectin) in AeC398 and AeV11 strains, and for Canavalia ensiformes (Con A, D-Man-binding lectin) in ATCC 15468, AeC398, AeQ32 and AeV11 strains, after bacterial growing at 37°C. On the other hand, specific receptors for WGA and EEL were completely abrogated growing the bacteria at 22°C. Binding studies with 125I- labeled lectins from WGA, EEL and Con A were performed. These assays essentially confirmed the selectivity, demonstrated in the agglutination assays of these lectins for the A. caviae strains. PMID:24031204

  5. Enantioselective carbohydrate recognition by synthetic lectins in water.

    PubMed

    Ríos, Pablo; Mooibroek, Tiddo J; Carter, Tom S; Williams, Christopher; Wilson, Miriam R; Crump, Matthew P; Davis, Anthony P

    2017-05-01

    Carbohydrate receptors with a chiral framework have been generated by combining a tetra-aminopyrene and a C3-symmetrical triamine via isophthalamide spacers bearing water-solubilising groups. These "synthetic lectins" are the first to show enantiodiscrimination in aqueous solution, binding N-acetylglucosamine (GlcNAc) with 16 : 1 enantioselectivity. They also show exceptional affinities. GlcNAc is bound with Ka up to 1280 M(-1), more than twice that measured for previous synthetic lectins, and three times the value for wheat germ agglutinin, the lectin traditionally employed to bind GlcNAc in glycobiological research. Glucose is bound with Ka = 250 M(-1), again higher than previous synthetic lectins. The results suggest that chirality can improve complementarity to carbohydrate substrates and may thus be advantageous in synthetic lectin design.

  6. Diversified Carbohydrate-Binding Lectins from Marine Resources

    PubMed Central

    Ogawa, Tomohisa; Watanabe, Mizuki; Naganuma, Takako; Muramoto, Koji

    2011-01-01

    Marine bioresources produce a great variety of specific and potent bioactive molecules including natural organic compounds such as fatty acids, polysaccharides, polyether, peptides, proteins, and enzymes. Lectins are also one of the promising candidates for useful therapeutic agents because they can recognize the specific carbohydrate structures such as proteoglycans, glycoproteins, and glycolipids, resulting in the regulation of various cells via glycoconjugates and their physiological and pathological phenomenon through the host-pathogen interactions and cell-cell communications. Here, we review the multiple lectins from marine resources including fishes and sea invertebrate in terms of their structure-activity relationships and molecular evolution. Especially, we focus on the unique structural properties and molecular evolution of C-type lectins, galectin, F-type lectin, and rhamnose-binding lectin families. PMID:22312473

  7. Recognition of microbial glycans by soluble human lectins.

    PubMed

    Wesener, Darryl A; Dugan, Amanda; Kiessling, Laura L

    2017-06-01

    Human innate immune lectins that recognize microbial glycans can conduct microbial surveillance and thereby help prevent infection. Structural analysis of soluble lectins has provided invaluable insight into how these proteins recognize their cognate carbohydrate ligands and how this recognition gives rise to biological function. In this opinion, we cover the structural features of lectins that allow them to mediate microbial recognition, highlighting examples from the collectin, Reg protein, galectin, pentraxin, ficolin and intelectin families. These analyses reveal how some lectins (e.g., human intelectin-1) can recognize glycan epitopes that are remarkably diverse, yet still differentiate between mammalian and microbial glycans. We additionally discuss strategies to identify lectins that recognize microbial glycans and highlight tools that facilitate these discovery efforts. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Lectins: an effective tool for screening of potential cancer biomarkers

    PubMed Central

    Jayapalan, Jaime Jacqueline; Lee, Cheng-Siang

    2017-01-01

    In recent years, the use of lectins for screening of potential biomarkers has gained increased importance in cancer research, given the development in glycobiology that highlights altered structural changes of glycans in cancer associated processes. Lectins, having the properties of recognizing specific carbohydrate moieties of glycoconjugates, have become an effective tool for detection of new cancer biomarkers in complex bodily fluids and tissues. The specificity of lectins provides an added advantage of selecting peptides that are differently glycosylated and aberrantly expressed in cancer patients, many of which are not possibly detected using conventional methods because of their low abundance in bodily fluids. When coupled with mass spectrometry, research utilizing lectins, which are mainly from plants and fungi, has led to identification of numerous potential cancer biomarkers that may be used in the future. This article reviews lectin-based methods that are commonly adopted in cancer biomarker discovery research. PMID:28894650

  9. Structure-function relationship of monocot mannose-binding lectins.

    PubMed Central

    Barre, A; Van Damme, E J; Peumans, W J; Rougé, P

    1996-01-01

    The monocot mannose-binding lectins are an extended superfamily of structurally and evolutionarily related proteins, which until now have been isolated from species of the Amaryllidaceae, Alliaceae, Araceae, Orchidaceae, and Liliaceae. To explain the obvious differences in biological activities, the structure-function relationships of the monocot mannose-binding lectins were studied by a combination of glycan-binding studies and molecular modeling using the deduced amino acid sequences of the currently known lectins. Molecular modeling indicated that the number of active mannose-binding sites per monomer varies between three and zero. Since the number of binding sites is fairly well correlated with the binding activity measured by surface plasmon resonance, and is also in good agreement with the results of previous studies of the biological activities of the mannose-binding lectins, molecular modeling is of great value for predicting which lectins are best suited for a particular application. PMID:8972598

  10. Monoclonal antibodies to endogenous galactose-specific tumor cell lectins.

    PubMed Central

    Raz, A; Meromsky, L; Carmi, P; Karakash, R; Lotan, D; Lotan, R

    1984-01-01

    A monoclonal antibody, 5D7, was obtained after immunization of syngeneic mice with B16 melanoma cell extracts enriched for endogenous lectin activity and screening for inhibition of lectin-mediated hemagglutination. Binding of this antibody to affinity-purified B16 melanoma galactoside-specific lectin was revealed by solid-phase radioimmunoassay and binding to the surface of viable B16 cells was demonstrated by indirect immunofluorescence. Inhibition of lectin activity and cell surface labeling by 5D7 antibody were also found with several types of cultured human and murine cells including melanoma, sarcoma and carcinoma. This monoclonal antibody should be useful for evaluating the role of tumor cell surface lectins in intercellular interactions and metastasis. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:6084591

  11. A developmentally regulated lectin in Bufo arenarum embryos.

    PubMed

    Elola, M T; Fink-de-Cabutti, N E; Herkovits, H

    1987-01-01

    We report the levels of an endogenous beta-galactoside lectin activity from Bufo arenarum whole embryos extracts and specific inhibition by saccharides at different developmental stages. Specific activity measured against trypsinized rabbit red blood cells showed relatively high and fluctuating levels during early stages (up to about 76 h post-fertilization) which fell to significantly lower and more constant values at late stages (77-264 h post-fertilization). Lactose is the most potent inhibitor of this lectin activity, and saccharides having alpha-galactoside configurations are weaker inhibitors. At the last embryonic stage, the agglutinating activity showed a different sugar specificity which suggests either the modification of the preexistent lectin or the synthesis of another type of lectin. The possible physiological roles of these lectins in the blockage of polyspermy or in embryonic cell-cell interactions are discussed.

  12. Ureaplasma urealyticum binds mannose-binding lectin.

    PubMed

    Benstein, Barbara D; Ourth, Donald D; Crouse, Dennis T; Shanklin, D Radford

    2004-10-01

    Mannose-binding C-type lectin (MBL) is an important component of innate immunity in mammals. Mannose-binding lectin (MBL), an acute phase protein, acts as an opsonin for phagocytosis and also activates the mannan-binding lectin complement pathway. It may play a particularly significant role during infancy before adequate specific protection can be provided by the adaptive immune system. Ureaplasma urealyticum has been linked to several diseases including pneumonia and chronic lung disease (CLD) in premature infants. We therefore investigated the ability of U. urealyticum to bind MBL. A guinea pig IgG anti-rabbit-MBL antiserum was produced. An immunoblot (dot-blot) assay done on nitrocellulose membrane determined that the anti-MBL antibody had specificity against both rabbit and human MBL. Pure cultures of U. urealyticum, serotype 3, were used to make slide preparations. The slides containing the organisms were then incubated with nonimmune rabbit serum containing MBL. Ureaplasma was shown to bind rabbit MBL with an immunocytochemical assay using the guinea pig IgG anti-rabbit MBL antiserum. Horseradish peroxidase (HRP)-labeled anti-guinea pig IgG was used to localize the reaction. The anti-MBL antiserum was also used in an immunocytochemical assay to localize U. urealyticum in histological sections of lungs from mice specifically infected with this organism. The same method also indicated binding of MBL by ureaplasma in human lung tissue obtained at autopsy from culture positive infants. Our results demonstrate that ureaplasma has the capacity to bind MBL. The absence of MBL may play a role in the predisposition of diseases related to this organism.

  13. MMBL proteins: from lectin to bacteriocin.

    PubMed

    Ghequire, Maarten G K; Loris, Remy; De Mot, René

    2012-12-01

    Arguably, bacteriocins deployed in warfare among related bacteria are among the most diverse proteinacous compounds with respect to structure and mode of action. Identification of the first prokaryotic member of the so-called MMBLs (monocot mannose-binding lectins) or GNA (Galanthus nivalis agglutinin) lectin family and discovery of its genus-specific killer activity in the Gram-negative bacteria Pseudomonas and Xanthomonas has added yet another kind of toxin to this group of allelopathic molecules. This novel feature is reminiscent of the protective function, on the basis of antifungal, insecticidal, nematicidal or antiviral activity, assigned to or proposed for several of the eukaryotic MMBL proteins that are ubiquitously distributed among monocot plants, but also occur in some other plants, fish, sponges, amoebae and fungi. Direct bactericidal activity can also be effected by a C-type lectin, but this is a mammalian protein that limits mucosal colonization by Gram-positive bacteria. The presence of two divergent MMBL domains in the novel bacteriocins raises questions about task distribution between modules and the possible role of carbohydrate binding in the specificity of target strain recognition and killing. Notably, bacteriocin activity was also demonstrated for a hybrid MMBL protein with an accessory protease-like domain. This association with one or more additional modules, often with predicted peptide-hydrolysing or -binding activity, suggests that additional bacteriotoxic proteins may be found among the diverse chimaeric MMBL proteins encoded in prokaryotic genomes. A phylogenetic survey of the bacterial MMBL modules reveals a mosaic pattern of strongly diverged sequences, mainly occurring in soil-dwelling and rhizosphere bacteria, which may reflect a trans-kingdom acquisition of the ancestral genes.

  14. Lectin activity in mycelial extracts of Fusarium species.

    PubMed

    Bhari, Ranjeeta; Kaur, Bhawanpreet; Singh, Ram S

    2016-01-01

    Lectins are non-immunogenic carbohydrate-recognizing proteins that bind to glycoproteins, glycolipids, or polysaccharides with high affinity and exhibit remarkable ability to agglutinate erythrocytes and other cells. In the present study, ten Fusarium species previously not explored for lectins were screened for the presence of lectin activity. Mycelial extracts of F. fujikuroi, F. beomiformii, F. begoniae, F. nisikadoi, F. anthophilum, F. incarnatum, and F. tabacinum manifested agglutination of rabbit erythrocytes. Neuraminidase treatment of rabbit erythrocytes increased lectin titers of F. nisikadoi and F. tabacinum extracts, whereas the protease treatment resulted in a significant decline in agglutination by most of the lectins. Results of hapten inhibition studies demonstrated unique carbohydrate specificity of Fusarium lectins toward O-acetyl sialic acids. Activity of the majority of Fusarium lectins exhibited binding affinity to d-ribose, l-fucose, d-glucose, l-arabinose, d-mannitol, d-galactosamine hydrochloride, d-galacturonic acid, N-acetyl-d-galactosamine, N-acetyl-neuraminic acid, 2-deoxy-d-ribose, fetuin, asialofetuin, and bovine submaxillary mucin. Melibiose and N-glycolyl neuraminic acid did not inhibit the activity of any of the Fusarium lectins. Mycelial extracts of F. begoniae, F. nisikadoi, F. anthophilum, and F. incarnatum interacted with most of the carbohydrates tested. F. fujikuroi and F. anthophilum extracts displayed strong interaction with starch. The expression of lectin activity as a function of culture age was investigated. Most species displayed lectin activity on the 7th day of cultivation, and it varied with progressing of culture age.

  15. Lectin binding to cystic stages of Taenia taeniaeformis.

    PubMed

    Sandeman, R M; Williams, J F

    1984-10-01

    Studies of membrane glycoconjugates of Taenia taeniaeformis were initiated by assays of the lectin binding characteristics of 35-day-old cysticerci. Parasites fixed in glutaraldehyde were incubated with one of the following FITC-labelled lectins: Concanavalin A (Con A), Lens culinaris agglutinin (LCA), Ricinus communis agglutinin (RCA), peanut agglutinin (PNA), fucose binding protein (FBP) and wheat germ agglutinin (WGA) and either their specific or a nonspecific sugar. Ultraviolet microscopy revealed that only Con A and LCA bound in large amounts to the surface of cysticerci. This binding was partly inhibited by the specific sugar, but the nonspecific sugar had little effect. The lectin not removed by either of the sugars may have been bound nonspecifically to the charged glycocalyx. Lectins were primarily bound on the anterior third of the parasite around the scolex invagination. Kinetic studies of lectin interactions were carried out with LCA and RCA by spectrophotofluorometric analysis of the amount bound specifically or nonspecifically over a range of lectin concentrations. Lens culinaris lectin binding was found to be specific and involve 2 receptors which showed large differences in their affinity for lectin and prevalence on the surface. Ricinus communis lectin did not bind specifically but nonspecific interactions were observed. Adherence of small numbers of host cells was shown to have no measurable effect on the lectin binding characteristics. The results suggest that the major surface carbohydrates exposed are D-mannose and/or D-glucose residues with the other sugar groups poorly represented. This relatively homogeneous surface may have implications for the antigenicity of the parasite in its host.

  16. Effects of Lectins on initial attachment of cariogenic Streptococcus mutans.

    PubMed

    Ito, Takashi; Yoshida, Yasuhiro; Shiota, Yasuyoshi; Ito, Yuki; Yamamoto, Tadashi; Takashiba, Shogo

    2017-09-08

    Oral bacteria initiate biofilm formation by attaching to tooth surfaces via an interaction of a lectin-like bacterial protein with carbohydrate chains on the pellicle. This study aimed to find naturally derived lectins that inhibit the initial attachment of a cariogenic bacterial species, Streptococcus mutans (S. mutans), to carbohydrate chains in saliva in vitro. Seventy kinds of lectins were screened for candidate motifs that inhibit the attachment of S. mutans ATCC 25175 to a saliva-coated culture plate. The inhibitory effect of the lectins on attachment of the S. mutans to the plates was quantified by crystal violet staining, and the biofilm was observed under a scanning electron microscope (SEM). Surface plasmon resonance (SPR) analysis was performed to examine the binding of S. mutans to carbohydrate chains and the binding of candidate lectins to carbohydrate chains, respectively. Moreover, binding assay between the biotinylated-lectins and the saliva components was conducted to measure the lectin binding. Lectins recognizing a salivary carbohydrate chain, Galβ1-3GalNAc, inhibited the binding of S. mutans to the plate. In particular, Agaricus bisporus agglutinin (ABA) markedly inhibited the binding. This inhibition was confirmed by SEM observation. SPR analysis indicated that S. mutans strongly binds to Galβ1-3GalNAc, and ABA binds to Galβ1-3GalNAc. Finally, the biotinylated Galβ1-3GalNAc-binding lectins including ABA demonstrated marked binding to the saliva components. These results suggest that ABA lectin inhibited the attachment of S. mutans to Galβ1-3GalNAc in saliva and ABA can be useful as a potent inhibitor for initial attachment of oral bacteria and biofilm formation.

  17. Isolation and characterization of lectins and lectin-alliinase complexes from bulbs of garlic (Allium sativum) and ramsons (Allium ursinum).

    PubMed

    Smeets, K; Van Damme, E J; Van Leuven, F; Peumans, W J

    1997-04-01

    A procedure developed to separate the homodimeric and heterodimeric mannose-binding lectins from bulbs of garlic (Allium sativum L.) and ramsons (Allium ursinum L.) also enabled the isolation of stable lectin-alliinase complexes. Characterization of the individual lectins indicated that, in spite of their different molecular structure, the homomeric and heteromeric lectins resemble each other reasonably well with respect to their agglutination properties and carbohydrate-binding specificity. However, a detailed analysis of the lectin-alliinase complexes from garlic and ramsons bulbs demonstrated that only the heterodimeric lectins are capable of binding to the glycan chains of the alliinase molecules (EC 4.4.1.4). Moreover, it appears that only a subpopulation of the alliinase molecules is involved in the formation of lectin-alliinase complexes and that the complexed alliinase contains more glycan chains than the free enzyme. Finally, some arguments are given that the lectin-alliinase complexes do not occur in vivo but are formed in vitro after homogenization of the tissue.

  18. The Liverwort Contains a Lectin That Is Structurally and Evolutionary Related to the Monocot Mannose-Binding Lectins1

    PubMed Central

    Peumans, Willy J.; Barre, Annick; Bras, Julien; Rougé, Pierre; Proost, Paul; Van Damme, Els J.M.

    2002-01-01

    A mannose (Man)-binding lectin has been isolated and characterized from the thallus of the liverwort Marchantia polymorpha. N-terminal sequencing indicated that the M. polymorpha agglutinin (Marpola) shares sequence similarity with the superfamily of monocot Man-binding lectins. Searches in the databases yielded expressed sequence tags encoding Marpola. Sequence analysis, molecular modeling, and docking experiments revealed striking structural similarities between Marpola and the monocot Man-binding lectins. Activity and specificity studies further indicated that Marpola is a much stronger agglutinin than the Galanthus nivalis agglutinin and exhibits a preference for methylated Man and glucose, which is unprecedented within the family of monocot Man-binding lectins. The discovery of Marpola allows us, for the first time, to corroborate the evolutionary relationship between a lectin from a lower plant and a well-established lectin family from flowering plants. In addition, the identification of Marpola sheds a new light on the molecular evolution of the superfamily of monocot Man-binding lectins. Beside evolutionary considerations, the occurrence of a G. nivalis agglutinin homolog in a lower plant necessitates the rethinking of the physiological role of the whole family of monocot Man-binding lectins. PMID:12114560

  19. Fluorescent carbohydrate probes for cell lectins

    NASA Astrophysics Data System (ADS)

    Galanina, Oxana; Feofanov, Alexei; Tuzikov, Alexander B.; Rapoport, Evgenia; Crocker, Paul R.; Grichine, Alexei; Egret-Charlier, Marguerite; Vigny, Paul; Le Pendu, Jacques; Bovin, Nicolai V.

    2001-09-01

    Fluorescein labeled carbohydrate (Glyc) probes were synthesized as analytical tools for the study of cellular lectins, i.e. SiaLe x-PAA-flu, Sia 2-PAA-flu, GlcNAc 2-PAA-flu, LacNAc-PAA-flu and a number of similar ones, with PAA a soluble polyacrylamide carrier. The binding of SiaLe x-PAA-flu was assessed using CHO cells transfected with E-selectin, and the binding of Sia 2-PAA-flu was assessed by COS cells transfected with siglec-9. In flow cytometry assays, the fluorescein probes demonstrated a specific binding to the lectin-transfected cells that was inhibited by unlabeled carbohydrate ligands. The intense binding of SiaLe x-PAA- 3H to the E-selectin transfected cells and the lack of binding to both native and permeabilized control cells lead to the conclusion that the polyacrylamide carrier itself and the spacer arm connecting the carbohydrate moiety with PAA did not contribute anymore to the binding. Tumors were obtained from nude mice by injection of CHO E-selectin or mock transfected cells. The fluorescent SiaLe x-PAA-flu probe could bind to the tumor sections from E-selectin positive CHO cells, but not from the control ones. Thus, these probes can be used to reveal specifically the carbohydrate binding sites on cells in culture as well as cells in tissue sections. The use of the confocal spectral imaging technique with Glyc-PAA-flu probes offered the unique possibility to detect lectins in different cells, even when the level of lectin expression was rather low. The confocal mode of spectrum recording provided an analysis of the probe localization with 3D submicron resolution. The spectral analysis (as a constituent part of the confocal spectral imaging technique) enabled interfering signals of the probe and intrinsic cellular fluorescence to be accurately separated, the distribution of the probe to be revealed and its local concentration to be measured.

  20. Studies on lectins. XXXVII. Isolation and characterization of the lectin from Jimson-weed seeds (Datura stramonium L.).

    PubMed

    Horejsí, V; Kocourek, J

    1978-01-25

    The lectin of Jimson-weed seeds (Datura stramonium L.) was isolated by affinity chromatography on a polysaccharide mixture from mycelium of Aspergillus niger. The lectin yields two bands on disc electrophoresis, it has sedimentation coefficient s20,w = 3.8 S and its apparent molecular weight estimated by thin layer gel chromatography is 120,000. The lectin reduced with mercaptoethanol yields on polyacrylamide gel electrophoresis in the presence of dodecyl sulfate three zones corresponding to subunits of molecular weight 72,000, 45,000 and 25,000. The lectin contains large amounts of cystine, glycine, 6.3% of hydroxyproline residues, 4.5% glucosamine and 28% of neutral sugar, predominantly arabinose. The lectin is nonspecific in human erythrocyte ABO system, it is not inhibited by simple sugars but is inhibited by a partial hydrolysate of chitin-containing mixture of polysaccharides from Aspergillus niger.

  1. Toshiaki Osawa: biochemistry of lectins and their applications in immunochemistry and cellular biology.

    PubMed

    Yamamoto, Kazuo; Irimura, Tatsuro

    2011-11-01

    Lectins are proteins that agglutinate cells and exhibit an antibody like, sugar-binding specificity. Professor Toshiaki Osawa has discovered, purified and characterized many plant lectins that display diverse biological activities. Using lectins as biochemical tools, he developed methods to determine the biochemical structures of glycoprotein glycans that react with lectins; separated and characterized glycoproteins and cell populations; analysed the mechanisms by which lectins activate cells; and characterized several cytokines produced by immune cells stimulated by lectins. The studies on lectins, the field he took strong leadership, developed into an essential hub of the biology of multicellular organisms.

  2. Bridging lectin binding sites by multivalent carbohydrates.

    PubMed

    Wittmann, Valentin; Pieters, Roland J

    2013-05-21

    Carbohydrate-protein interactions are involved in a multitude of biological recognition processes. Since individual protein-carbohydrate interactions are usually weak, multivalency is often required to achieve biologically relevant binding affinities and selectivities. Among the possible mechanisms responsible for binding enhancement by multivalency, the simultaneous attachment of a multivalent ligand to several binding sites of a multivalent receptor (i.e. chelation) has been proven to have a strong impact. This article summarizes recent examples of chelating lectin ligands of different size. Covered lectins include the Shiga-like toxin, where the shortest distance between binding sites is ca. 9 Å, wheat germ agglutinin (WGA) (shortest distance between binding sites 13-14 Å), LecA from Pseudomonas aeruginosa (shortest distance 26 Å), cholera toxin and heat-labile enterotoxin (shortest distance 31 Å), anti-HIV antibody 2G12 (shortest distance 31 Å), concanavalin A (ConA) (shortest distance 72 Å), RCA120 (shortest distance 100 Å), and Erythrina cristagalli (ECL) (shortest distance 100 Å). While chelating binding of the discussed ligands is likely, experimental proof, for example by X-ray crystallography, is limited to only a few cases.

  3. [Separation of osteoclasts by lectin affinity chromatography].

    PubMed

    Itokazu, M; Tan, A; Tanaka, S

    1991-09-01

    Newborn rat calvaria bone cells obtained by digestion were fractionated on columns of wheat-germ agglutinin (WGA) sepharose 6MB for osteoclast isolation. The initial nonspecific binding cells which were passed through the WGA sepharose column by a buffer acquired a high enzyme activity of alkaline phosphatase, but not that of acid phosphatase. However, elution of cells using a buffer with the addition of N-acetyl-D-glucosamine resulted in a high acid phosphatase activity but no alkaline phosphatase activity. The former WGA binding negative fraction enriched osteoblasts averaging 30 microns in size. The latter WGA binding positive fraction enriched osteoclasts ranging from 20 microns to 60 microns in size. The electron-microscope clearly demonstrated the cellular details of osteoclasts. Isolated cell counts showed a ratio of six to four. These results indicate that our method of osteoclast isolation is simple and useful in lectin affinity chromatography because all cells have sugar moieties on their surface and the binding of osteoclasts can be reversed by the addition of specific lectin-binding sugars to the eluting buffer.

  4. Glycoprotein enrichment through lectin affinity techniques.

    PubMed

    Mechref, Yehia; Madera, Milan; Novotny, Milos V

    2008-01-01

    Posttranslational modifications (PTM) of proteins are among the key biological regulators of function, activity, localization, and interaction. The fact that no more than 30,000-50,000 proteins are encoded by the human genome underlines the importance of posttranslational modifications in modulating the activities and functions of proteins in health and disease. With approximately 50% of all proteins now considered to be glycosylated, its physiological importance in mammalian systems is imperative. Aberrant glycosylation has now been recognized as an attribute of many mammalian diseases, including hereditary disorders, immune deficiencies, neurodegenerative diseases, cardiovascular conditions, and cancer. As many potential disease biomarkers may be glycoproteins present in only minute quantities in tissue extracts and physiological fluids, glycoprotein isolation and enrichment may be critical in a search for such biomarkers. For decades, efforts have been focused on the development of glycoprotein enrichment from complex biological samples. Logically, the great majority of these enrichment methodologies rely on the use of immobilized lectins, which permit selective enrichment of the pools of glycoproteins for proteomic/glycomic studies. In this chapter, lectin affinity chromatography in different formats are described, including tubes; packed columns, and microfluidic channels.

  5. Lectin domains at the frontiers of plant defense.

    PubMed

    Lannoo, Nausicaä; Van Damme, Els J M

    2014-01-01

    Plants are under constant attack from pathogens and herbivorous insects. To protect and defend themselves, plants evolved a multi-layered surveillance system, known as the innate immune system. Plants sense their encounters upon perception of conserved microbial structures and damage-associated patterns using cell-surface and intracellular immune receptors. Plant lectins and proteins with one or more lectin domains represent a major part of these receptors. The whole group of plant lectins comprises an elaborate collection of proteins capable of recognizing and interacting with specific carbohydrate structures, either originating from the invading organisms or from damaged plant cell wall structures. Due to the vast diversity in protein structures, carbohydrate recognition domains and glycan binding specificities, plant lectins constitute a very diverse protein superfamily. In the last decade, new types of nucleocytoplasmic plant lectins have been identified and characterized, in particular lectins expressed inside the nucleus and the cytoplasm of plant cells often as part of a specific plant response upon exposure to different stress factors or changing environmental conditions. In this review, we provide an overview on plant lectin motifs used in the constant battle against pathogens and predators during plant defenses.

  6. Use of labeled tomato lectin for imaging vasculature structures.

    PubMed

    Robertson, Richard T; Levine, Samantha T; Haynes, Sherry M; Gutierrez, Paula; Baratta, Janie L; Tan, Zhiqun; Longmuir, Kenneth J

    2015-02-01

    Intravascular injections of fluorescent or biotinylated tomato lectin were tested to study labeling of vascular elements in laboratory mice. Injections of Lycopersicon esculentum agglutinin (tomato lectin) (50-100 µg/100 µl) were made intravascularly, through the tail vein, through a cannula implanted in the jugular vein, or directly into the left ventricle of the heart. Tissues cut for thin 10- to 12-µm cryostat sections, or thick 50- to 100-µm vibratome sections, were examined using fluorescence microscopy. Tissue labeled by biotinylated lectin was examined by bright field microscopy or electron microscopy after tissue processing for biotin. Intravascular injections of tomato lectin led to labeling of vascular structures in a variety of tissues, including brain, kidney, liver, intestine, spleen, skin, skeletal and cardiac muscle, and experimental tumors. Analyses of fluorescence in serum indicated the lectin was cleared from circulating blood within 2 min. Capillary labeling was apparent in tissues collected from animals within 1 min of intravascular injections, remained robust for about 1 h, and then declined markedly until difficult to detect 12 h after injection. Light microscopic images suggest the lectin bound to the endothelial cells that form capillaries and endothelial cells that line some larger vessels. Electron microscopic studies confirmed the labeling of luminal surfaces of endothelial cells. Vascular labeling by tomato lectin is compatible with a variety of other morphological labeling techniques, including histochemistry and immunocytochemistry, and thus appears to be a sensitive and useful method to reveal vascular patterns in relationship to other aspects of parenchymal development, structure, and function.

  7. Extensive amino acid sequence homologies between animal lectins

    SciTech Connect

    Paroutaud, P.; Levi, G.; Teichberg, V.I.; Strosberg, A.D.

    1987-09-01

    The authors have established the amino acid sequence of the ..beta..-D-galactoside binding lectin from the electric eel and the sequences of several peptides from a similar lectin isolated from human placenta. These sequences were compared with the published sequences of peptides derived from the ..beta..-D-galactoside binding lectin from human lung and with sequences deduced from cDNAs assigned to the ..beta..-D-galactoside binding lectins from chicken embryo skin and human hepatomas. Significant homologies were observed. One of the highly conserved regions that contains a tryptophan residue and two glutamic acid resides is probably part of the ..beta..-D-galactoside binding site, which, on the basis of spectroscopic studies of the electric eel lectin, is expected to contain such residues. The similarity of the hydropathy profiles and the predicted secondary structure of the lectins from chicken skin and electric eel, in spite of differences in their amino acid sequences, strongly suggests that these proteins have maintained structural homologies during evolution and together with the other ..beta..-D-galactoside binding lectins were derived form a common ancestor gene.

  8. Lectins, agglutinins, and their roles in autoimmune reactivities.

    PubMed

    Vojdani, Aristo

    2015-01-01

    Lectins are carbohydrate-binding proteins present throughout nature that act as agglutinins. Approximately 30% of our food contains lectins, some of which may be resistant enough to digestion to enter the circulation. Because of their binding properties, lectins can cause nutrient deficiencies, disrupt digestion, and cause severe intestinal damage when consumed in excess by an individual with dysfunctional enzymes. These effects are followed by disruption of intestinal barrier integrity, which is the gateway to various autoimmunities. Shared amino acid motifs between dietary lectins, exogenous peptides, and various body tissues may lead to cross-reactivity, resulting in the production of antibodies against lectin and bacterial antigens, followed by autoimmunity. The detection of immunoglobulin G (IgG) or immunoglobulin A (IgA) antibodies against specific lectins may serve as a guide for the elimination of these lectins from the diet. It is proposed that this process can reduce the peripheral antigenic stimulus and, thereby, result in a diminution of disease symptoms in some-but not all-patients with autoimmune disorders.

  9. Lectins: getting familiar with translators of the sugar code.

    PubMed

    André, Sabine; Kaltner, Herbert; Manning, Joachim C; Murphy, Paul V; Gabius, Hans-Joachim

    2015-01-22

    The view on the significance of the presence of glycans in glycoconjugates is undergoing a paradigmatic change. Initially mostly considered to be rather inert and passive, the concept of the sugar code identifies glycans as highly versatile platform to store information. Their chemical properties endow carbohydrates to form oligomers with unsurpassed structural variability. Owing to their capacity to engage in hydrogen (and coordination) bonding and C-H/π-interactions these "code words" can be "read" (in Latin, legere) by specific receptors. A distinct class of carbohydrate-binding proteins are the lectins. More than a dozen protein folds have developed carbohydrate-binding capacity in vertebrates. Taking galectins as an example, distinct expression patterns are traced. The availability of labeled endogenous lectins facilitates monitoring of tissue reactivity, extending the scope of lectin histochemistry beyond that which traditionally involved plant lectins. Presentation of glycan and its cognate lectin can be orchestrated, making a glycan-based effector pathway in growth control of tumor and activated T cells possible. In order to unravel the structural basis of lectin specificity for particular glycoconjugates mimetics of branched glycans and programmable models of cell surfaces are being developed by strategic combination of lectin research with synthetic and supramolecular chemistry.

  10. Lectin domains at the frontiers of plant defense

    PubMed Central

    Lannoo, Nausicaä; Van Damme, Els J. M.

    2014-01-01

    Plants are under constant attack from pathogens and herbivorous insects. To protect and defend themselves, plants evolved a multi-layered surveillance system, known as the innate immune system. Plants sense their encounters upon perception of conserved microbial structures and damage-associated patterns using cell-surface and intracellular immune receptors. Plant lectins and proteins with one or more lectin domains represent a major part of these receptors. The whole group of plant lectins comprises an elaborate collection of proteins capable of recognizing and interacting with specific carbohydrate structures, either originating from the invading organisms or from damaged plant cell wall structures. Due to the vast diversity in protein structures, carbohydrate recognition domains and glycan binding specificities, plant lectins constitute a very diverse protein superfamily. In the last decade, new types of nucleocytoplasmic plant lectins have been identified and characterized, in particular lectins expressed inside the nucleus and the cytoplasm of plant cells often as part of a specific plant response upon exposure to different stress factors or changing environmental conditions. In this review, we provide an overview on plant lectin motifs used in the constant battle against pathogens and predators during plant defenses. PMID:25165467

  11. Lessons learned from mice deficient in lectin complement pathway molecules.

    PubMed

    Genster, Ninette; Takahashi, Minoru; Sekine, Hideharu; Endo, Yuichi; Garred, Peter; Fujita, Teizo

    2014-10-01

    The lectin pathway of the complement system is initiated when the pattern-recognition molecules, mannose-binding lectin (MBL), ficolins or collectin-11, bind to invading pathogens or damaged host cells. This leads to activation of MBL/ficolin/collectin-11 associated serine proteases (MASPs), which in turn activate downstream complement components, ultimately leading to elimination of the pathogen. Mice deficient in the key molecules of lectin pathway of complement have been generated in order to build knowledge of the molecular mechanisms of the lectin pathway in health and disease. Despite differences in the genetic arrangements of murine and human orthologues of lectin pathway molecules, the knockout mice have proven to be valuable models to explore the effect of deficiency states in humans. In addition, new insight and unexpected findings on the diverse roles of lectin pathway molecules in complement activation, pathogen infection, coagulation, host tissue injury and developmental biology have been revealed by in vivo investigations. This review provides an overview of the mice deficient in lectin pathway molecules and highlights some of the most important findings that have resulted from studies of these. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Cloning and characterization of root-specific barley lectin

    SciTech Connect

    Lerner, D.R.; Raikhel, N.V. )

    1989-09-01

    Cereal lectins are a class of biochemically and antigenically related proteins localized in a tissue-specific manner in embryos and adult plants. To study the specificity of lectin expression, a barley (Hordeum vulgare L.) embryo cDNa library was constructed and a clone (BLc3) for barley lectin was isolated. BLc3 is 972 nucleotides long and includes an open reading frame of 212 amino acids. The deduced amino acid sequence contains a putative signal peptide of 26 amino acid residues followed by a 186 amino acid polypeptide. This polypeptide has 95% sequence identity to the antigenically indistinguishable wheat germ agglutinin isolectin-B (WGA-B) suggesting that BLc3 encodes barley lectin. Further evidence that BLc3 encodes barley lectin was obtained by immunoprecipitation of the in vitro translation products of BLc3 RNA transcripts and barley embryo poly(A{sup +}) RNA. In situ hybridizations with BLc3 showed that barley lectin gene expression is confined to the outermost cell layers of both embryonic and adult root tips. On Northern blots, BLc3 hybridizes to a 1.0 kilobyte mRNA in poly(A{sup +}) RNA from both embryos and root tips. We suggest, on the basis of immunoblot experiments, that barley lectin is synthesized as a glycosylated precursor and processed by removal of a portion of the carboxyl terminus including the single N-linked glycosylation site.

  13. Cloning and Characterization of Root-Specific Barley Lectin 1

    PubMed Central

    Lerner, David R.; Raikhel, Natasha V.

    1989-01-01

    Cereal lectins are a class of biochemically and antigenically related proteins localized in a tissue-specific manner in embryos and adult plants. To study the specificity of lectin expression, a barley (Hordeum vulgare L.) embryo cDNA library was constructed and a clone (BLc3) for barley lectin was isolated. BLc3 is 972 nucleotides long and includes an open reading frame of 212 amino acids. The deduced amino acid sequence contains a putative signal peptide of 26 amino acid residues followed by a 186 amino acid polypeptide. This polypeptide has 95% sequence identity to the antigenically indistinguishable wheat germ agglutinin isolectin-B (WGA-B) suggesting that BLc3 encodes barley lectin. Further evidence that BLc3 encodes barley lectin was obtained by immunoprecipitation of the in vitro translation products of BLc3 RNA transcripts and barley embryo poly(A+) RNA. In situ hybridizations with BLc3 showed that barley lectin gene expression is confined to the outermost cell layers of both embryonic and adult root tips. On Northern blots, BLc3 hybridizes to a 1.0 kilobyte mRNA in poly(A+) RNA from both embryos and root tips. We suggest, on the basis of immunoblot experiments, that barley lectin is synthesized as a glycosylated precursor and processed by removal of a portion of the carboxyl terminus including the single N-linked glycosylation site. Images Figure 2 Figure 3 Figure 4 Figure 5 PMID:16666982

  14. Purification, some properties of a D-galactose-binding leaf lectin from Erythrina indica and further characterization of seed lectin.

    PubMed

    Konozy, Emadeldin H E; Mulay, Ranjana; Faca, Vitor; Ward, Richard John; Greene, Lewis Joel; Roque-Barriera, Maria Cristina; Sabharwal, Sushma; Bhide, Shobhana V

    2002-10-01

    Lectin from a leaf of Erythrina indica was isolated by affinity chromatography on Lactamyl-Seralose 4B. Lectin gave a single band in polyacrylamide gel electrophoresis (PAGE). In SDS-gel electrophoresis under reducing and non-reducing conditions Erythrina indica leaf lectin (EiLL) split into two bands with subunit molecular weights of 30 and 33 kDa, whereas 58 kDa was obtained for the intact lectin by gel filtration on Sephadex G-100. EiLL agglutinated all human RBC types, with a slight preference for the O blood group. Lectin was found to be a glycoprotein with a neutral sugar content of 9.5%. The carbohydrate specificity of lectin was directed towards D-galactose and its derivatives with pronounced preference for lactose. EiLL had pH optima at pH 7.0; above and below this pH lectin lost sugar-binding capability rapidly. Lectin showed broad temperature optima from 25 to 50 degrees C; however, at 55 degrees C EiLL lost more than 90% of its activity and at 60 degrees C it was totally inactivated. The pI of EiLL was found to be 7.6. The amino acid analysis of EiLL indicated that the lectin was rich in acidic as well as hydrophobic amino acids and totally lacked cysteine and methionine. The N-terminal amino acids were Val-Glu-Thr-IIe-Ser-Phe-Ser-Phe-Ser-Glu-Phe-Glu-Ala-Gly-Asn-Asp-X-Leu-Thr-Gln-Glu-Gly-Ala-Ala-Leu-. Chemical modification studies of both EiLL and Erythrina indica seed lectin (EiSL) with phenylglyoxal, DEP and DTNB revealed an absence of arginine, histidine and cysteine, respectively, in or near the ligand-binding site of both lectins. Modification of tyrosine with NAI led to partial inactivation of EiLL and EiSL; however, total inactivation was observed upon NBS-modification of two tryptophan residues in EiSL. Despite the apparent importance of these tryptophan residues for lectin activity they did not seem to have a direct role in binding haptenic sugar as D-galactose did not protect lectin from inactivation by NBS.

  15. [Current knowledge about presumable functions of plant lectins].

    PubMed

    Shakirova, F M; Bezrukova, M V

    2007-01-01

    Current data on the diversity of plant lectins and their functional importance for plants, caused primarily by their capacity to link carbohydrate ligands specifically and convertibly, are reviewed. For instance, the role of plant lectins in the recognition of alien organisms and in the adaptation of plants to various stress-induced effects is discussed. In addition to centres of specific affinity to carbohydrates, plant lectins are characterized by the presence of sites responsible for hydrophobic interactions with non-carbohydrate molecules. These sites link to plant hormones, proteins, and other metabolites, thus participating in the regulation of metabolic processes controlling growth, development, and differentiation in plants. The structure and biological properties of ribosome-inactivating proteins having and not having lectin activity are discussed, as well as their role in plant protection from pests and pathogens. Current data on the assumed functions of the independent groups of plant lectins with specific endogenic role are given. These include chitin-specific lectins synthesized in phloem, which are capable of forming protein-protein and RNA-protein complexes and translocating via vessels, which thus play their specific intra- or intercellular interactions, processes of growth, development, and protection of plants. Other groups of plant lectins, induced by jasmonate, such as Nictaba (Nicotiana tabaccum agglutinin), and cereal lectins related to jacalin, which are localised in the cytoplasm and nucleus, probably play regulatory role in the formation of stress response in plants. The structure and currently discussed functions of wheat germ agglutinin, a typical representative of cereal lectins, are analysed in detail.

  16. Self-assembled carbohydrate-based vesicles for lectin targeting.

    PubMed

    Dos Santos, Marinalva Cardoso; Micheletto, Yasmine Miguel Serafini; da Silveira, Nadya Pesce; da Silva Pinto, Luciano; Giacomelli, Fernando Carlos; de Lima, Vânia Rodrigues; Frizon, Tiago Elias Allievi; Dal-Bó, Alexandre Gonçalves

    2016-12-01

    This study examined the physicochemical interactions between vesicles formed by phosphatidylcholine (PC) and glycosylated polymeric amphiphile N-acetyl-β-d-glucosaminyl-PEG900-docosanate (C22PEG900GlcNAc) conjugated with Bauhinia variegata lectin (BVL). Lectins are proteins or glycoproteins capable of binding glycosylated membrane components. Accordingly, the surface functionalization by such entities is considered a potential strategy for targeted drug delivery. We observed increased hydrodynamic radii (RH) of PC+C22PEG900GlcNAc vesicles in the presence of lectins, suggesting that this aggregation was due to the interaction between lectins and the vesicular glycosylated surfaces. Furthermore, changes in the zeta potential of the vesicles with increasing lectin concentrations implied that the vesicular glycosylated surfaces were recognized by the investigated lectin. The presence of carbohydrate residues on vesicle surfaces and the ability of the vesicles to establish specific interactions with BVL were further explored using atomic force microscopy (AFM) and small-angle X-ray scattering (SAXS) analysis. The results indicated that the thickness of the hydrophilic layer was to some extent influenced by the presence of lectins. The presence of lectins required a higher degree of polydispersity as indicated by the width parameter of the log-normal distribution of size, which also suggested more irregular structures. Reflectance Fourier transform infrared (HATR-FTIR), differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis.) analyses revealed that the studied lectin preferentially interacted with the choline and carbonyl groups of the lipid, thereby changing the choline orientation and intermolecular interactions. The protein also discretely reduced the intermolecular communication of the hydrophobic acyl chains, resulting in a disordered state.

  17. Generation of fibronectin receptors on macrophages by wheat germ lectin.

    PubMed

    Hörmann, H; Jelinić, V; Richter, H

    1983-08-01

    A chymotrypsin-derived and 125I-labelled 125-kDa fragment of human plasma fibronectin which contained the cell binding site, was only weakly bound by peritoneal macrophages of guinea pigs and binding was not saturable. In presence of wheat germ lectin binding increased proportionally to the logarithm of the lectin concentration. Association of 125I-fragment with cells was partially prevented by non-labelled fragment indicating a saturable receptor-ligand interaction. An apparent affinity constant of about 2--4 x 10(-5) M was evaluated. A considerable fraction of the cell-bound 125I-fragment resisted removal by proteases suggesting that it was internalized. In order to investigate an influence of wheat germ lectin on the binding of 125I-fibronectin by the cells the macrophages were preincubated with the lectin followed by washing and evaluation of 125I-fibronectin binding. A simultaneous incubation of the cells with 125I-fibronectin and lectin was impractical due to partial interaction of the two proteins giving rise to some unspecific precipitates. Preincubation with wheat germ lectin considerably improved the capacity of the macrophages for binding of 125I-fibronectin. Again the binding of 125I-labelled protein could be restricted by unlabelled one. N-acetyl-glucosamine inhibited the binding of 125I-fibronectin by wheat germ lectin-treated cells if applied in the preincubation phase and more effectively, if applied in the final 125I-fibronectin binding assay. N-Acetylneuraminic acid also inhibited this step. In addition to wheat germ lectin concanavalin A was capable of generating fibronectin receptors on the cell surface. Soy bean lectin, however, was ineffective.

  18. Fine specificities of two lectins from Cymbosema roseum seeds: a lectin specific for high-mannose oligosaccharides and a lectin specific for blood group H type II trisaccharide.

    PubMed

    Dam, Tarun K; Cavada, Benildo S; Nagano, Celso S; Rocha, Bruno Am; Benevides, Raquel G; Nascimento, Kyria S; de Sousa, Luiz Ag; Oscarson, Stefan; Brewer, C Fred

    2011-07-01

    The legume species of Cymbosema roseum of Diocleinae subtribe produce at least two different seed lectins. The present study demonstrates that C. roseum lectin I (CRL I) binds with high affinity to the "core" trimannoside of N-linked oligosaccharides. Cymbosema roseum lectin II (CRL II), on the other hand, binds with high affinity to the blood group H trisaccharide (Fucα1,2Galα1-4GlcNAc-). Thermodynamic and hemagglutination inhibition studies reveal the fine binding specificities of the two lectins. Data obtained with a complete set of monodeoxy analogs of the core trimannoside indicate that CRL I recognizes the 3-, 4- and 6-hydroxyl groups of the α(1,6) Man residue, the 3- and 4-hydroxyl group of the α(1,3) Man residue and the 2- and 4-hydroxyl groups of the central Man residue of the trimannoside. CRL I possesses enhanced affinities for the Man5 oligomannose glycan and a biantennary complex glycan as well as glycoproteins containing high-mannose glycans. On the other hand, CRL II distinguishes the blood group H type II epitope from the Lewis(x), Lewis(y), Lewis(a) and Lewis(b) epitopes. CRL II also distinguishes between blood group H type II and type I trisaccharides. CRL I and CRL II, respectively, possess differences in fine specificities when compared with other reported mannose and fucose recognizing lectins. This is the first report of a mannose-specific lectin (CRL I) and a blood group H type II-specific lectin (CRL II) from seeds of a member of the Diocleinae subtribe.

  19. Lectins as inducers of interferon-gamma production in human lymphocytes: lentil lectin is highly efficient.

    PubMed

    Rönnblom, L; Funa, K; Ersson, B; Alm, G V

    1982-10-01

    Several of many tested plant lectins induced interferon (IFN) production in cultures of human peripheral blood mononuclear leucocytes (PBL). The mannose-binding lectin obtained from Lens culinaris (LCL) was a particularly efficient inducer of trypsin-sensitive antiviral activity, which qualified as IFN-gamma because it was 90-95% destroyed by pH 2 treatment but not neutralized by anti-IFN-gamma antibodies. However, such antibodies neutralized the residual 5-10% pH 2-resistant IFN, which therefore represented IFN-alpha. Further evidence for the IFN-gamma nature of the LCL-induced IFN was that its production in PBL cultures required both T lymphocytes and macrophages and that its induction of antiviral resistance in human amnion cells was significantly delayed compared with IFN-alpha. Under optimal conditions LCL induced titres of IFN-gamma corresponding to more than 20,000 IFN-alpha units/ml medium, higher than observed with other tested, established IFN-gamma inducers. Other desirable properties of this lectin, as discussed, also suggest that it will be of value for efficient large-scale IFN-gamma production.

  20. [Galectins, a class of unconventional lectins].

    PubMed

    Advedissian, Tamara; Deshayes, Frédérique; Poirier, Françoise; Grandjean, Cyrille; Viguier, Mireille

    2015-05-01

    Galectins constitute a family of soluble animal lectins defined by their evolutionary conserved carbohydrate recognition domain and their affinity for β-galactosides containing glycoconjugates. Each galectin is characterized by a specific spatio-temporal distribution and a unique set of ligands and molecular partners. Interestingly, galectins are found both extracellularly and intracellularly and modulate various cellular processes. Knock-out mutant mice for galectins-1, 3 or 7 are viable but display a wide range of defects under various stress conditions. Indeed, galectins are multifunctional proteins involved in cell-cell and cell-extracellular matrix interactions, organization of membrane domains, cell signalling and also in intracellular trafficking, apoptosis, regulation of cell cycle. Galectins represent potential therapeutic targets, especially in the context of cancer and inflammatory diseases. © 2015 médecine/sciences – Inserm.

  1. Isolation and characterization of a lectin from Annona muricata seeds.

    PubMed

    Damico, D C S; Freire, M G M; Gomes, V M; Toyama, M H; Marangoni, S; Novello, J C; Macedo, M L R

    2003-11-01

    A lectin with a high affinity for glucose/mannose was isolated from Annona muricata seeds (Annonaceae) by gel filtration chromatography on Sephacryl S-200, ion exchange chromatography on a DEAE SP-5 PW column, and molecular exclusion on a Protein Pak Glass 300 SW column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (PAGE) yielded two protein bands of approximately 14 kDa and 22 kDa. However, only one band was seen in native PAGE. The Mr of the lectin estimated by fast-performance liquid chromatography-gel filtration on Superdex 75 was 22 kDa. The lectin was a glycoprotein with 8% carbohydrate (neutral sugar) and required divalent metal cations (Ca2+, Mg2+, and Mn2+) for full activity. Amino acid analysis revealed a large content of Glx, Gly, Phe, and Lys. The lectin agglutinated dog, chicken, horse, goose, and human erythrocytes and inhibited the growth of the fungi Fusarium oxysporum, Fusarium solani, and Colletotrichum musae.

  2. Lectins stain cells differentially in the coral, Montipora capitata

    USGS Publications Warehouse

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  3. Structure and Function of Mammalian Carbohydrate-Lectin Interactions

    NASA Astrophysics Data System (ADS)

    Anderson, Kevin; Evers, David; Rice, Kevin G.

    Over the past three decades the field of glycobiology has expanded beyond a basic understanding of the structure and biosynthesis of glycoprotein, proteoglycans, and glycolipids toward a more detailed picture of how these molecules afford communication through binding to mammalian lectins. Although the number of different mammalian lectin domains appears to be finite and even much smaller than early estimates predicated based on the diversity of glycan structures, nature appears capable of using these in numerous combinations to fine tune specificity. The following provides an overview of the major classes of mammalian lectins and discusses their glycan binding specificity. The review provides a snapshot of the field of glycobiology that continues to grow providing an increasing number of examples of biological processes that rely upon glycan-lectin binding.

  4. Sweet complementarity: the functional pairing of glycans with lectins.

    PubMed

    Gabius, H-J; Manning, J C; Kopitz, J; André, S; Kaltner, H

    2016-05-01

    Carbohydrates establish the third alphabet of life. As part of cellular glycoconjugates, the glycans generate a multitude of signals in a minimum of space. The presence of distinct glycotopes and the glycome diversity are mapped by sugar receptors (antibodies and lectins). Endogenous (tissue) lectins can read the sugar-encoded information and translate it into functional aspects of cell sociology. Illustrated by instructive examples, each glycan has its own ligand properties. Lectins with different folds can converge to target the same epitope, while intrafamily diversification enables functional cooperation and antagonism. The emerging evidence for the concept of a network calls for a detailed fingerprinting. Due to the high degree of plasticity and dynamics of the display of genes for lectins the validity of extrapolations between different organisms of the phylogenetic tree yet is inevitably limited.

  5. An alternate high yielding purification method for Clitoria ternatea lectin.

    PubMed

    Naeem, Aabgeena; Ahmad, Ejaz; Khan, Rizwan Hasan

    2007-10-01

    In our previous publication we had reported the purification and characterization of Clitoria ternatea agglutinin from its seeds on fetuin CL agarose affinity column, designated CTA [A. Naeem, S. Haque, R.H. Khan. Protein J., 2007]. Since CTA binds beta-d-galactosides, this lectin can be used as valuable tool for glycobiology studies in biomedical and cancer research. So an attempt was made for a high yielding alternative purification method employing the use of asialofetuin CL agarose column for the above-mentioned lectin, designated CTL. The fetuin affinity purified agglutinin was found similar to asialofetuin affinity purified lectin in SDS pattern, HPLC and N-terminal sequence. The content of lectin was found to be 30mg/30g dry weight of pulse. The yield was 2.8% as compared to 0.3% obtained on fetuin column. The number of tryptophan and tyrosine estimated was four and six per subunit.

  6. Molecular cloning of mannose-binding lectins from Clivia miniata.

    PubMed

    Van Damme, E J; Smeets, K; Van Leuven, F; Peumans, W J

    1994-03-01

    Screening of a cDNA library constructed from total RNA isolated from young developing ovaries of Clivia miniata Regel with the amaryllis lectin cDNA clone resulted in the isolation of four different isolectin clones which clearly differ from each other in their nucleotide sequences and hence also in their deduced amino acid sequences. Apparently the lectin is translated from an mRNA of ca. 800 nucleotides encoding a precursor polypeptide of 163 amino acids. Northern blot analysis of total RNA isolated from different tissues of Clivia miniata has shown that the lectin is expressed in most plant tissues with very high lectin mRNA concentrations in the ovary and the seed endosperm.

  7. Insights into animal and plant lectins with antimicrobial activities.

    PubMed

    Dias, Renata de Oliveira; Machado, Leandro Dos Santos; Migliolo, Ludovico; Franco, Octavio Luiz

    2015-01-05

    Lectins are multivalent proteins with the ability to recognize and bind diverse carbohydrate structures. The glyco -binding and diverse molecular structures observed in these protein classes make them a large and heterogeneous group with a wide range of biological activities in microorganisms, animals and plants. Lectins from plants and animals are commonly used in direct defense against pathogens and in immune regulation. This review focuses on sources of animal and plant lectins, describing their functional classification and tridimensional structures, relating these properties with biotechnological purposes, including antimicrobial activities. In summary, this work focuses on structural-functional elucidation of diverse lectin groups, shedding some light on host-pathogen interactions; it also examines their emergence as biotechnological tools through gene manipulation and development of new drugs.

  8. Specific interaction of lectins with liposomes and monolayers bearing neoglycolipids.

    PubMed

    Faivre, Vincent; Costa, Maria de Lourdes; Boullanger, Paul; Baszkin, Adam; Rosilio, Véronique

    2003-10-01

    The interaction of three lectins (wheat germ, Ulex europaeus I, and Lotus tetragonolobus agglutinins: WGA, UEA-I and LTA) with either N-acetyl-D-glucosamine or L-fucose neoglycolipids incorporated into phospholipid monolayers and liposome bilayers was studied at the air/water interface and in bulk solution. The results show that for both systems studied, synthesized neoglycolipids were capable of binding their specific lectin and that, in general, the binding of lectins increased with the increase in the molar fraction of the saccharide derivative incorporated in either the monolayers or bilayers. However, whereas for UEA-I, molecular recognition was enhanced by a strong hydrophobic interaction, for WGA and LTA successful recognition was predominantly related to the distance between neighboring sugar groups. The observed lengthy adsorption times of these lectins onto their specific ligands were attributed to interfacial conformational changes occurring in the proteins upon their adsorption at the interfaces.

  9. A Lectin from Dioclea violacea Interacts with Midgut Surface of Lutzomyia migonei, Unlike Its Homologues, Cratylia floribunda Lectin and Canavalia gladiata Lectin

    PubMed Central

    Monteiro Tínel, Juliana Montezuma Barbosa; Benevides, Melina Fechine Costa; Frutuoso, Mércia Sindeaux; Rocha, Camila Farias; Arruda, Francisco Vassiliepe Sousa; Vasconcelos, Mayron Alves; Pereira-Junior, Francisco Nascimento; Cajazeiras, João Batista; do Nascimento, Kyria Santiago; Martins, Jorge Luiz; Teixeira, Edson Holanda; Cavada, Benildo Sousa; dos Santos, Ricardo Pires; Lima Pompeu, Margarida Maria

    2014-01-01

    Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand fly. Susceptibility and refractoriness to Leishmania depend on the outcome of multiple interactions that take place within the sand fly gut. Promastigote attachment to sand fly midgut epithelium is essential to avoid being excreted together with the digested blood meal. Promastigote and gut sand fly surface glycans are important ligands in this attachment. The purpose of the present study was to evaluate the interaction of three lectins isolated from leguminous seeds (Diocleinae subtribe), D-glucose and D-mannose-binding, with glycans on Lutzomyia migonei midgut. To study this interaction the lectins were labeled with FITC and a fluorescence assay was performed. The results showed that only Dioclea violacea lectin (DVL) was able to interact with midgut glycans, unlike Cratylia floribunda lectin (CFL) and Canavalia gladiata lectin (CGL). Furthermore, when DVL was blocked with D-mannose the interaction was inhibited. Differences of spatial arrangement of residues and volume of carbohydrate recognition domain (CRD) may be the cause of the fine specificity of DVL for glycans in the surface on Lu. migonei midgut. The findings in this study showed the presence of glycans in the midgut with glucose/mannose residues in its composition and these residues may be important in interaction between Lu. migonei midgut and Leishmania. PMID:25431778

  10. A lectin from Dioclea violacea Interacts with midgut surface of Lutzomyia migonei, unlike its homologues, Cratylia floribunda lectin and Canavalia gladiata lectin.

    PubMed

    Monteiro Tínel, Juliana Montezuma Barbosa; Benevides, Melina Fechine Costa; Frutuoso, Mércia Sindeaux; Rocha, Camila Farias; Arruda, Francisco Vassiliepe Sousa; Vasconcelos, Mayron Alves; Pereira-Junior, Francisco Nascimento; Cajazeiras, João Batista; do Nascimento, Kyria Santiago; Martins, Jorge Luiz; Teixeira, Edson Holanda; Cavada, Benildo Sousa; dos Santos, Ricardo Pires; Lima Pompeu, Margarida Maria

    2014-01-01

    Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand fly. Susceptibility and refractoriness to Leishmania depend on the outcome of multiple interactions that take place within the sand fly gut. Promastigote attachment to sand fly midgut epithelium is essential to avoid being excreted together with the digested blood meal. Promastigote and gut sand fly surface glycans are important ligands in this attachment. The purpose of the present study was to evaluate the interaction of three lectins isolated from leguminous seeds (Diocleinae subtribe), D-glucose and D-mannose-binding, with glycans on Lutzomyia migonei midgut. To study this interaction the lectins were labeled with FITC and a fluorescence assay was performed. The results showed that only Dioclea violacea lectin (DVL) was able to interact with midgut glycans, unlike Cratylia floribunda lectin (CFL) and Canavalia gladiata lectin (CGL). Furthermore, when DVL was blocked with D-mannose the interaction was inhibited. Differences of spatial arrangement of residues and volume of carbohydrate recognition domain (CRD) may be the cause of the fine specificity of DVL for glycans in the surface on Lu. migonei midgut. The findings in this study showed the presence of glycans in the midgut with glucose/mannose residues in its composition and these residues may be important in interaction between Lu. migonei midgut and Leishmania.

  11. Interactions between Rhizobia and Lectins of Lentil, Pea, Broad Bean, and Jackbean 1

    PubMed Central

    Wong, Peter P.

    1980-01-01

    A quantitative method was developed to measure the binding of fluorescent-labeled lentil (Lens esculenta Moench), pea (Pisum sativum L.), broad bean (Vicia faba L.), and jackbean (Canavalia ensiformis L., DC.) lectins to various Rhizobium strains. Lentil lectin bound to three of the five Rhizobium leguminosarum strains tested. The number of lentil lectin molecules bound per R. leguminosarum 128C53 cell was 2.1 × 104. Lentil lectin also bound to R. japonicum 61A133. Pea and broad bean lectins bound to only two of the five strains of R. leguminosarum, whereas concanavalin A (jackbean lectin) bound to all strains of R. leguminosarum, R. phaseoli, R. japonicum, and R. sp. tested. Since these four lectins have similar sugarbinding properties but different physical properties, the variation in bindings of these lectins to various Rhizobium strains indicates that binding of lectin to Rhizobium is determined not only by the sugar specificity of the lectin but also by its physical characteristics. The binding of lentil lectin and concanavalin A to R. leguminosarum 128C53 could be inhibited by glucose, fructose, and mannose. However, even at 150 millimolar glucose, about 15% of the binding remained. The binding of lentil lectin to R. japonicum 61A133 could be inhibited by glucose but not by galactose. It is concluded that the binding site of lentil lectin to R. japonicum is different from the binding site of soybean lectin to R. japonicum. PMID:16661328

  12. A lectin with some unique characteristics from the samta tomato.

    PubMed

    Wang, H; Ng, T B

    2006-04-01

    A lectin, with a molecular mass of 79 kDa, and with specificity toward rhamnose and O-nitrophenyl-beta-D-galactopyranoside, was isolated from samta tomato fruits. The procedure entailed ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose and gel filtration by fast protein liquid chromatography on Superdex 75. The lectin was unadsorbed on DEAE-cellulose but adsorbed on Affi-gel blue gel and CM-cellulose. The lectin was stable up to 70 degrees C. The lectin activity was potentiated by NaOH solutions (25-100 mM), but was reduced by 50 and 100 mM HCl solutions. The activity of the lectin was reduced in the presence of CaCl(2), MgCl(2) and ZnCl(2), but was potentiated by 5 and 10 mM AlCl(3) solutions. The lectin stimulated the mitogenic response in mouse splenocytes and inhibited human immunodeficiency virus-1 reverse transcriptase with an IC(50) of 6.2 microM.

  13. Structural analysis of Centrolobium tomentosum seed lectin with inflammatory activity.

    PubMed

    Almeida, Alysson Chaves; Osterne, Vinicius Jose da Silva; Santiago, Mayara Queiroz; Pinto-Junior, Vanir Reis; Silva-Filho, Jose Caetano; Lossio, Claudia Figueiredo; Nascimento, Francisco Lucas Faustino; Almeida, Ricardo Patricio Honorato; Teixeira, Claudener Souza; Leal, Rodrigo Bainy; Delatorre, Plinio; Rocha, Bruno Anderson Matias; Assreuy, Ana Maria Sampaio; Nascimento, Kyria Santiago; Cavada, Benildo Sousa

    2016-04-15

    A glycosylated lectin (CTL) with specificity for mannose and glucose has been detected and purified from seeds of Centrolobium tomentosum, a legume plant from Dalbergieae tribe. It was isolated by mannose-sepharose affinity chromatography. The primary structure was determined by tandem mass spectrometry and consists of 245 amino acids, similar to other Dalbergieae lectins. CTL structures were solved from two crystal forms, a monoclinic and a tetragonal, diffracted at 2.25 and 1.9 Å, respectively. The carbohydrate recognition domain (CRD), metal-binding site and glycosylation site were characterized, and the structural basis for mannose/glucose-binding was elucidated. The lectin adopts the canonical dimeric organization of legume lectins. CTL showed acute inflammatory effect in paw edema model. The protein was subjected to ligand screening (dimannosides and trimannoside) by molecular docking, and interactions were compared with similar lectins possessing the same ligand specificity. This is the first crystal structure of mannose/glucose native seed lectin with proinflammatory activity isolated from the Centrolobium genus. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Vasorelaxant activity of Canavalia grandiflora seed lectin: A structural analysis.

    PubMed

    Barroso-Neto, Ito Liberato; Simões, Rafael Conceição; Rocha, Bruno Anderson Matias; Bezerra, Maria Julia Barbosa; Pereira-Junior, Francisco Nascimento; Silva Osterne, Vinicius José; Nascimento, Kyria Santiago; Nagano, Celso Shiniti; Delatorre, Plinio; Pereira, Maria Gonçalves; Freitas Pires, Alana; Sampaio, Alexandre Holanda; Assreuy, Ana Maria Sampaio; Cavada, Benildo Sousa

    2014-02-01

    Lectins are comprised of a large family of proteins capable of the specific and reversible recognition of carbohydrates. Legume lectins, the most studied plant lectins, show high structural similarity, but with modifications that imply a variation in the intensity of some biological activities. In this work, the primary and tertiary structures of Canavalia grandiflora (ConGF) were determined. ConGF, a lectin isolated from C. grandiflora seeds, is able to induce relaxant activity in rat aortic rings. The complete sequence of ConGF comprises 237 amino acids. This particular protein has primary sequence variations commonly found in lectins from Dioclea and Canavalia genera. The protein structure was solved at 2.3 Å resolution by X-ray crystallography. An X-Man molecule was modeled into the carbohydrate recognition domain. Still, ConGF (30 and 100 μg mL(-1)) elicited 25% of vasorelaxation (IC50=34.48 ± 5.07 μg mL(-1)) in endothelialized aortic rings. A nonselective inhibitor of nitric oxide blocked ConGF relaxant effect, showing mediation by nitric oxide. Key distances between ConGF carbohydrate recognition domain residues were determined in order to explain this effect, in turn revealing some structural aspects that could differentiate lectins from the Canavalia genera with respect to different efficacy in vasorelaxant effect. Copyright © 2013 Elsevier Inc. All rights reserved.

  15. Antifungal activity of lectins against yeast of vaginal secretion

    PubMed Central

    Gomes, Bruno Severo; Siqueira, Ana Beatriz Sotero; de Cássia Carvalho Maia, Rita; Giampaoli, Viviana; Teixeira, Edson Holanda; Arruda, Francisco Vassiliepe Sousa; do Nascimento, Kyria Santiago; de Lima, Adriana Nunes; Souza-Motta, Cristina Maria; Cavada, Benildo Sousa; Porto, Ana Lúcia Figueiredo

    2012-01-01

    Lectins are carbohydrate-binding proteins of non-imune origin. This group of proteins is distributed widely in nature and they have been found in viruses, microorganisms, plants and animals. Lectins of plants have been isolated and characterized according to their chemical, physical-chemical, structural and biological properties. Among their biological activities, we can stress its fungicidal action. It has been previously described the effect of the lectins Dviol, DRL, ConBr and LSL obtained from the seeds of leguminous plants on the growth of yeasts isolated from vaginal secretions. In the present work the experiments were carried out in microtiter plates and the results interpreted by both methods: visual observations and a microplate reader at 530nm. The lectin concentrations varied from 0.5 to 256μg/mL, and the inoculum was established between 65-70% of trammitance. All yeast samples isolated from vaginal secretion were evaluated taxonomically, where were observed macroscopic and microscopic characteristics to each species. The LSL lectin did not demonstrate any antifungal activity to any isolate studied. The other lectins DRL, ConBr and DvioL, showed antifungal potential against yeast isolated from vaginal secretion. These findings offering offer a promising field of investigation to develop new therapeutic strategies against vaginal yeast infections, collaborating to improve women's health. PMID:24031889

  16. Glycoprotein targeting and other applications of lectins in biotechnology.

    PubMed

    Naeem, Aabgeena; Saleemuddin, M; Khan, Rizwan Hasan

    2007-06-01

    Glycoconjugates comprise a variety of structures, include glycoproteins and glycolipids and are found on the surfaces of animal and plant cells, as well as on the surface of microorganisms. Determination of the structure and the distribution of glycoconjugates on cell surfaces are important for the understanding their biological function. Lectins are useful to investigate protein-carbohydrate interactions, because they have specificity for defined carbohydrate structure. They have been implicated in cell-to-cell recognition and signaling, blood group typing, in immune recognition process, and various other biological processes, such as viral, bacterial, mycoplasmal and parasitic infections, fertilization, cancer metastasis, growth and differentiation. Once thought to be confined to plant seeds, lectins are now recognized as ubiquitous in virtually all living systems, ranging from viruses and bacteria to animals. Plant lectins provide a rich source of carbohydrate-recognizing protein reagents for glycobiologists and biotechnologists. Biotechnology offers the therapeutic use of lectin against certain life threatening diseases such as human immunodeficiency virus and cancer. This review presents a comprehensive summary of research efforts that focus on the actual and potential uses and advantages of using lectins to target glycoproteins and also glycoproteins to target lectins.

  17. Purification and cDNA cloning of a lectin and a lectin-like protein from Apios americana Medikus tubers.

    PubMed

    Kouzuma, Yoshiaki; Irie, Satoshi; Yamazaki, Rikiya; Yonekura, Masami

    2014-01-01

    An Apios americana lectin (AAL) and a lectin-like protein (AALP) were purified from tubers by chromatography on Butyl-Cellulofine, ovomucoid-Cellulofine, and DEAE-Cellulofine columns. AAL showed strong hemagglutinating activity toward chicken and goose erythrocytes, but AALP showed no such activity toward any of the erythrocytes tested. The hemagglutinating activity of AAL was not inhibited by mono- or disaccharides, but was inhibited by glycoproteins, such as asialofetuin and ovomucoid, suggesting that AAL is an oligosaccharide-specific lectin. The cDNAs of AAL and AALP consist of 1,093 and 1,104 nucleotides and encode proteins of 302 and 274 amino acid residues, respectively. Both amino acid sequences showed high similarity to known legume lectins, and those of their amino acids involved in carbohydrate and metal binding were conserved.

  18. Structures and binding specificity of galactose- and mannose-binding lectins from champedak: differences from jackfruit lectins.

    PubMed

    Gabrielsen, Mads; Abdul-Rahman, Puteri Shafinaz; Othman, Shatrah; Hashim, Onn H; Cogdell, Richard J

    2014-06-01

    Galactose-binding and mannose-binding lectins from the champedak fruit, which is native to South-east Asia, exhibit useful potential clinical applications. The specificity of the two lectins for their respective ligands allows the detection of potential cancer biomarkers and monitoring of the glycosylated state of proteins in human serum and/or urine. To fully understand and expand the use of these natural proteins, their complete sequences and crystal structures are presented here, together with details of sugar binding.

  19. Utilization of lectin-histochemistry in forensic neuropathology: lectin staining provides useful information for postmortem diagnosis in forensic neuropathology.

    PubMed

    Nishi, Katsuji; Tanegashima, Akio; Yamamoto, Yoshio; Ushiyama, Ikuko; Ikemoto, Keiko; Yamasaki, Shigeru; Nishimura, Akiyoshi; Rand, Steven; Brinkmann, Bernd

    2003-09-01

    We have investigated the deposition of glycoconjugates in human brain tissue with or without brain disorders. In this review we describe the application of lectin-histochemistry techniques to forensic neuropathology. Lectin staining is able to reveal several kinds of carbohydrate-related depositions in addition to the conventional degenerative changes including senile plaques, neurofibrillary tangles and corpora amylacea. The senile plaques and neurofibrillary tangles were clearly stained by Con A, PSA and GSI lectins, the corpora amylacea which is relevant to repeated brain hypoxia and mitochondrial damage was also easily detected by these and many other kinds of lectins. Amorphous spaces were detected around blood vessels and independently from blood vessels by lectin staining in the white matter from patients with brain disorders or severe edema. The white matter lesions were not considered relevant for forensic pathology, until a large group of cerebral white matter lesions were detected in the elderly with increasing frequency by modern neuro-imaging methods. The spherical deposits were newly detected by lectin staining in the molecular layer of the dentate gyrus of the hippocampal formation chiefly from patients with schizophrenia or cognitive dysfunctions.

  20. Lectin-like molecules in transcriptome of Littorina littorea hemocytes.

    PubMed

    Gorbushin, Alexander M; Borisova, Elena A

    2015-01-01

    The common periwinkle Littorina littorea was introduced in the list of models for comparative immunobiology as a representative of phylogenetically important taxon Caenogastropoda. Using Illumina sequencing technology, we de novo assembled the transcriptome of Littorina littorea hemocytes from 182 million mRNA-Seq pair-end 100 bp reads into a total of 15,526 contigs clustered in 4472 unigenes. The transcriptome profile was analyzed for presence of carbohydrate-binding molecules in a variety of architectural contexts. Hemocytes' repertoire of lectin-like proteins bearing conserved carbohydrate-recognition domains (CRDs) is highly diversified, including 11 of 15 lectin families earlier described in animals, as well as the novel members of lectin family found for the first time in mollusc species. The new molluscan lineage-specific domain combinations were confirmed by cloning and sequencing, including the fuco-lectin related molecules (FLReMs) composed of N-terminal region with no sequence homology to any known protein, a middle Fucolectin Tachylectin-4 Pentaxrin (FTP) domain, and a C-terminal epidermal growth factor (EGF) repeat region. The repertoire of lectin-like molecules is discussed in terms of their potential participation in the receptor phase of immune response. In total, immune-associated functions may be attributed to 70 transcripts belonging to 6 lectin families. These lectin-like genes show low overlap between species of invertebrates, suggesting relatively rapid evolution of immune-associated genes in the group. The repertoire provides valuable candidates for further characterization of the gene functions in mollusc immunity.

  1. Genome-wide identification and domain organization of lectin domains in cucumber.

    PubMed

    Dang, Liuyi; Van Damme, Els J M

    2016-11-01

    Lectins are ubiquitous proteins in plants and play important roles in a diverse set of biological processes, such as plant defense and cell signaling. Despite the availability of the Cucumis sativus L. genome sequence since 2009, little is known with respect to the occurrence of lectins in cucumber. In this study, a total of 146 putative lectin genes belonging to 10 different lectin families were identified and localized in the cucumber genome. Domain architecture analysis revealed that most of these lectin gene sequences contain multiple domains, where lectin domains are linked with other domains, as such creating chimeric lectin sequences encoding proteins with dual activities. This study provides an overview of lectin motifs in cucumber and will help to understand their potential biological role(s). Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  2. Structure predictions of two Bauhinia variegata lectins reveal patterns of C-terminal properties in single chain legume lectins.

    PubMed

    Moreira, Gustavo M S G; Conceição, Fabricio R; McBride, Alan J A; Pinto, Luciano da S

    2013-01-01

    Bauhinia variegata lectins (BVL-I and BVL-II) are single chain lectins isolated from the plant Bauhinia variegata. Single chain lectins undergo post-translational processing on its N-terminal and C-terminal regions, which determines their physiological targeting, carbohydrate binding activity and pattern of quaternary association. These two lectins are isoforms, BVL-I being highly glycosylated, and thus far, it has not been possible to determine their structures. The present study used prediction and validation algorithms to elucidate the likely structures of BVL-I and -II. The program Bhageerath-H was chosen from among three different structure prediction programs due to its better overall reliability. In order to predict the C-terminal region cleavage sites, other lectins known to have this modification were analysed and three rules were created: (1) the first amino acid of the excised peptide is small or hydrophobic; (2) the cleavage occurs after an acid, polar, or hydrophobic residue, but not after a basic one; and (3) the cleavage spot is located 5-8 residues after a conserved Leu amino acid. These rules predicted that BVL-I and -II would have fifteen C-terminal residues cleaved, and this was confirmed experimentally by Edman degradation sequencing of BVL-I. Furthermore, the C-terminal analyses predicted that only BVL-II underwent α-helical folding in this region, similar to that seen in SBA and DBL. Conversely, BVL-I and -II contained four conserved regions of a GS-I association, providing evidence of a previously undescribed X4+unusual oligomerisation between the truncated BVL-I and the intact BVL-II. This is the first report on the structural analysis of lectins from Bauhinia spp. and therefore is important for the characterisation C-terminal cleavage and patterns of quaternary association of single chain lectins.

  3. Structure Predictions of Two Bauhinia variegata Lectins Reveal Patterns of C-Terminal Properties in Single Chain Legume Lectins

    PubMed Central

    Moreira, Gustavo M. S. G.; Conceição, Fabricio R.; McBride, Alan J. A.; Pinto, Luciano da S.

    2013-01-01

    Bauhinia variegata lectins (BVL-I and BVL-II) are single chain lectins isolated from the plant Bauhinia variegata. Single chain lectins undergo post-translational processing on its N-terminal and C-terminal regions, which determines their physiological targeting, carbohydrate binding activity and pattern of quaternary association. These two lectins are isoforms, BVL-I being highly glycosylated, and thus far, it has not been possible to determine their structures. The present study used prediction and validation algorithms to elucidate the likely structures of BVL-I and -II. The program Bhageerath-H was chosen from among three different structure prediction programs due to its better overall reliability. In order to predict the C-terminal region cleavage sites, other lectins known to have this modification were analysed and three rules were created: (1) the first amino acid of the excised peptide is small or hydrophobic; (2) the cleavage occurs after an acid, polar, or hydrophobic residue, but not after a basic one; and (3) the cleavage spot is located 5-8 residues after a conserved Leu amino acid. These rules predicted that BVL-I and –II would have fifteen C-terminal residues cleaved, and this was confirmed experimentally by Edman degradation sequencing of BVL-I. Furthermore, the C-terminal analyses predicted that only BVL-II underwent α-helical folding in this region, similar to that seen in SBA and DBL. Conversely, BVL-I and -II contained four conserved regions of a GS-I association, providing evidence of a previously undescribed X4+unusual oligomerisation between the truncated BVL-I and the intact BVL-II. This is the first report on the structural analysis of lectins from Bauhinia spp. and therefore is important for the characterisation C-terminal cleavage and patterns of quaternary association of single chain lectins. PMID:24260572

  4. Probing the cons and pros of lectin-induced immunomodulation: case studies for the mistletoe lectin and galectin-1.

    PubMed

    Gabius, H J

    2001-07-01

    When imagining to monitor animal cells through a microscope with resolution at the molecular level, a salient attribute of their surfaces will be the abundance of glycan chains. They present galactosides at their termini widely extending like tentacles into the extracellular space. Their spatial accessibility and their potential for structural variability endow especially these glycan parts with capacity to act as docking points for molecular sensors (sugar receptors such as lectins). Binding and ligand clustering account for transmission of post-binding signals into the cell interior. The range of triggered activities has turned plant lectins into popular tools in cell biology and immunology. Potential for clinical application has been investigated rigorously only in recent years. As documented in vitro and in vivo for the galactoside-specific mistletoe lectin, its apparent immunomodulatory capacity reflected in upregulation of production of proinflammatory cytokines will not necessarily be clinically favorable but a double-edged sword. In fact, lectin application has been shown to stimulate tumor growth in cell lines, histocultures of human tumors and in two animal models using chemical carcinogenesis or tumor transplantation. When testing immunological effects of the endogenous lectin galectin-1, protection against disorders mediated by activated T cells came up for consideration. Elimination of these cells via CD7-dependent induction of apoptosis, and a shift to the Th2 response by the galectin, are factors to ameliorate disease states. This result encourages further efforts with other galectins. Functional redundancy, synergism, diversity or antagonism among galectins are being explored to understand the actual role of this class of endogenous lectins in inflammation. Regardless of the results of further preclinical testing for galectin-1, these two case studies break new ground in our understanding how glycans as ligands for lectins convey reactivity to

  5. The production and properties of an antiserum to potato (Solanum tuberosum) lectin.

    PubMed Central

    Ashford, D; Allen, A K; Neuberger, A

    1982-01-01

    Precipitation of potato (Solanum tuberosum) lectin by antisera was not affected by treatments that abolish lectin activity. An antiserum precipitated glycosylated derivatives of the lectin but not a deglycosylated peptide. The haemagglutination inhibition titre of this antiserum was not affected by removing anti-glycopeptide antibodies. This evidence suggests that the antiserum contains two populations of antibodies, specific for different domains of the lectin. Images Fig. 1. Fig. 2. PMID:6807287

  6. Bauhinia variegata var. variegata lectin: isolation, characterization, and comparison.

    PubMed

    Chan, Yau Sang; Ng, Tzi Bun

    2015-01-01

    Bauhinia variegata var. variegata seeds are rich in proteins. Previously, one of the major storage proteins of the seeds was found to be a trypsin inhibitor that possessed various biological activities. By using another purification protocol, a glucoside- and galactoside-binding lectin that demonstrated some differences from the previously reported B. variegata lectin could be isolated from the seeds. It involved affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Q-Sepharose and Mono Q, and also size exclusion chromatography on Superdex 75. The lectin was not retained on Affi-gel blue gel but interacted with Q-Sepharose. The lectin was a 64-kDa protein with two 32-kDa subunits. It had low thermostability (stable up to 50 °C) and moderate pH stability (stable in pH 3-10). It exhibited anti-proliferative activity on nasopharyngeal carcinoma HONE1 cells with an IC50 of 12.8 μM after treatment for 48 h. It also slightly inhibited the growth of hepatoma HepG2 cells. The lectin may have potential in aiding cancer treatments.

  7. Molecular biology of seed storage proteins and lectins

    SciTech Connect

    Shannon, L.M.; Chrispeels, M.J.

    1986-01-01

    The seeds of many plants contain abundant storage protein and lectin which have been the subject of biochemical investigations for over a hundred years. Because these proteins represent abundant gene products translated from abundant messages, they were among the first plant proteins to which the techniques of molecular biology were applied. Many of the proteins have now been purified and characterized and their amino acid sequences have been determined; some have been crystallized and their 3-dimensional structure is known. Studies of their biosynthesis, transport and accumulation in protein bodies have lead to a greater understanding of the dynamics of plant cell organelles. Seed storage proteins and lectins are encoded by small gene families whose members show considerable homology and appear to have been conserved in evolution. The expression of these genes is highly regulated in time (development) and space (tissue). The recent discovery that other plant organs synthesize lectins, or lectin-like proteins which are closely related to the seed lectins, has lent additional support to the search for a function for these intriguing proteins; this finding also indicates that different members of the gene family are expressed in different tissues. Individual papers were processed separately for the data base.

  8. Prevalence of the F-type lectin domain.

    PubMed

    Bishnoi, Ritika; Khatri, Indu; Subramanian, Srikrishna; Ramya, T N C

    2015-08-01

    F-type lectins are fucolectins with characteristic fucose and calcium-binding sequence motifs and a unique lectin fold (the "F-type" fold). F-type lectins are phylogenetically widespread with selective distribution. Several eukaryotic F-type lectins have been biochemically and structurally characterized, and the F-type lectin domain (FLD) has also been studied in the bacterial proteins, Streptococcus mitis lectinolysin and Streptococcus pneumoniae SP2159. However, there is little knowledge about the extent of occurrence of FLDs and their domain organization, especially, in bacteria. We have now mined the extensive genomic sequence information available in the public databases with sensitive sequence search techniques in order to exhaustively survey prokaryotic and eukaryotic FLDs. We report 437 FLD sequence clusters (clustered at 80% sequence identity) from eukaryotic, eubacterial and viral proteins. Domain architectures are diverse but mostly conserved in closely related organisms, and domain organizations of bacterial FLD-containing proteins are very different from their eukaryotic counterparts, suggesting unique specialization of FLDs to suit different requirements. Several atypical phylogenetic associations hint at lateral transfer. Among eukaryotes, we observe an expansion of FLDs in terms of occurrence and domain organization diversity in the taxa Mollusca, Hemichordata and Branchiostomi, perhaps coinciding with greater emphasis on innate immune strategies in these organisms. The naturally occurring FLDs with diverse domain organizations that we have identified here will be useful for future studies aimed at creating designer molecular platforms for directing desired biological activities to fucosylated glycoconjugates in target niches.

  9. Lectin Activity in Gut Extract of Culex pipiens

    PubMed Central

    Koosha, Mona; Abai, Mohammad Reza; Abolhasani, Mandan; Charedar, Soroor; Basseri, Hamid Reza

    2013-01-01

    Background: The role of lectins is important in interaction between pathogens and mosquito vectors. This study was performed to identify agglutinin activities of protein molecules on the midgut of Culex pipiens. Methods: Culex pipiens was reared in insectray condition and the midguts of males and females (blood fed and unfed) were dissected separately in Tris-HCl buffer. The extracts of midguts were applied for hemagglutinin assay against red blood cells of rabbit, mouse, rat, dog, horse, sheep, guinea pig, cow, human (A, B, AB, O groups). Then, the RBCs with relatively high agglutinin activity were chosen for carbohydrate inhibition assay. D (+) glucose, D (+) galactose, D (+) mannose, D (−) fructose, D (−) arabinose, L (−) fucose, lactose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, sialic acid were used to specify carbohydrate binding lectin. Results: The highest agglutinin activities were found against sheep and rabbits RBCs. Sexual diversity of agglutinin activities was observed among midgut extraction of males and females. In addition, variation in agglutinin activity of blood fed and unfed female mosquitoes were detected. The lectin activity was inhibited highly with glucose, galactose, fucose and fructose but less inhibitor activities was observed by arabinose, N-acetyl-D-galactosamine, n-acetyl-d-glucosamine, lactose and mannose. Conclusion: The secretion of hemagglutinins (lectins or lectin-like molecules) in the digestive system depends on the type of food in the gut. This suggests that emptying of the gut in preparation for protein rich food probably starts the secretion of hemagglutinins. PMID:23785692

  10. Recombinant lectins: an array of tailor-made glycan-interaction biosynthetic tools.

    PubMed

    Oliveira, Carla; Teixeira, José A; Domingues, Lucília

    2013-03-01

    Lectins are a heterogeneous group of proteins found in plants, animals and microorganisms, which possess at least one non-catalytic domain that binds reversibly to specific mono- or oligosaccharides. The range of lectins and respective biological activities is unsurprising given the immense diversity and complexity of glycan structures and the multiple modes of interaction with proteins. Recombinant DNA technology has been traditionally used for cloning and characterizing newly discovered lectins. It has also been employed as a means of producing pure and sequence-defined lectins for different biotechnological applications. This review focuses on the production of recombinant lectins in heterologous organisms, and highlighting the Escherichia coli and Pichia pastoris expression systems, which are the most employed. The choice of expression host depends on the lectin. Non-glycosylated recombinant lectins are produced in E. coli and post-translational modified recombinant lectins are produced in eukaryotic organisms, namely P. pastoris and non-microbial hosts such as mammalian cells. Emphasis is given to the applications of the recombinant lectins especially (a) in cancer diagnosis and/or therapeutics, (b) as anti-microbial, anti-viral, and anti-insect molecules or (c) in microarrays for glycome profiling. Most reported applications are from recombinant plant lectins. These applications benefit from the tailor-made design associated with recombinant production and will aid in unraveling the complex biological mechanisms of glycan-interactions, bringing recombinant lectins to the forefront of glycobiology. In conclusion, recombinant lectins are developing into valuable biosynthetic tools for biomedical research.

  11. Histological and lectin histochemical studies on the olfactory and respiratory mucosae of the sheep.

    PubMed

    Ibrahim, Dalia; Nakamuta, Nobuaki; Taniguchi, Kazumi; Yamamoto, Yoshio; Taniguchi, Kazuyuki

    2014-03-01

    The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL), Datura stramonium lectin (DSL), Soybean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-120), Erythrina cristagalli lectin (ECL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L). The associated glands of the olfactory mucosa, Bowman's glands, were stained with 13 lectins. While both the goblet cells and mucous nasal glands were stained with 8 lectins; five of them (WGA, s-WGA, STL, Vicia villosa agglutinin (VVA) and ECL) were mutually positive among the Bowman's glands, mucous nasal glands and the goblet cells. These findings indicate that the glycohistochemical characteristics of the free borders of both olfactory and respiratory epithelia are similar to each other, suggesting that secretions from the Bowman's glands and those of the goblet cells and mucous nasal glands are partially exchanged between the surface of two epithelia to contribute the functions of the respiratory epithelium and the olfactory receptor cells, respectively.

  12. Studies on lectins. XLIX. The use of glycosyl derivatives of Dextran T-500 for affinity electrophoresis of lectins.

    PubMed

    Cerovský, V; Tichá, M; Horejsi, V; Kocourek, J

    1980-09-01

    p-Aminophenyl glycosides and glycosylamines were coupled to periodate oxidized Dextran T-500 either directly or through an epsilon-aminocaproic acid spacer. The new glycosylated derivatives of dextran specifically precipitate lectins having the appropriate carbohydrate specificity, and thus were used in the preparation of affinity gels for affinity electrophoresis of lectins. The apparent strength of interaction of several lectins with carbohydrate residues immobilized in this way was less than with carbohydrates immobilized in O-glycosyl polyacrylamide copolymers. The presence of epsilon-aminocaproic spacer had no effect on the strength of interaction. The advantages of this type of macromolecular derivative of the ligand for affinity electrophoresis and some differences between the glycosylated dextrans and O-glycosyl polyacrylamide copolymers are discussed. Dextrans containing bound p-aminophenyl alpha-D-mannopyranoside and p-aminophenyl alpha-D-glucopyranoside were used to study the binding properties of concanavalin A and the lectin from Lathyrus sativus seeds. For the investigation of interaction of lectins from Ricinus communis and Glycine soja seeds, dextran derivatives containing bound p-aminophenyl alpha- and beta-D-galactopyranosides and alpha- and beta-D-galactopyranosylamines were used.

  13. Marine sponge lectins: actual status on properties and biological activities.

    PubMed

    Gomes Filho, Sandro Mascena; Cardoso, Juscélio Donizete; Anaya, Katya; Silva do Nascimento, Edilza; de Lacerda, José Thalles Jucelino Gomes; Mioso, Roberto; Santi Gadelha, Tatiane; de Almeida Gadelha, Carlos Alberto

    2014-12-26

    Marine sponges are primitive metazoans that produce a wide variety of molecules that protect them against predators. In studies that search for bioactive molecules, these marine invertebrates stand out as promising sources of new biologically-active molecules, many of which are still unknown or little studied; thus being an unexplored biotechnological resource of high added value. Among these molecules, lectins are proteins that reversibly bind to carbohydrates without modifying them. In this review, various structural features and biological activities of lectins derived from marine sponges so far described in the scientific literature are discussed. From the results found in the literature, it could be concluded that lectins derived from marine sponges are structurally diverse proteins with great potential for application in the production of biopharmaceuticals, especially as antibacterial and antitumor agents.

  14. Symbiotic Bacteria Direct Expression of an Intestinal Bactericidal Lectin

    PubMed Central

    Cash, Heather L.; Whitham, Cecilia V.; Behrendt, Cassie L.; Hooper, Lora V.

    2009-01-01

    The mammalian intestine harbors complex societies of beneficial bacteria that are maintained in the lumen with minimal penetration of mucosal surfaces. Microbial colonization of germ-free mice triggers epithelial expression of RegIIIγ, a secreted C-type lectin. RegIIIγ binds intestinal bacteria but lacks the complement recruitment domains present in other microbe-binding mammalian C-type lectins. We show that RegIIIγ and its human counterpart, HIP/PAP, are directly antimicrobial proteins that bind their bacterial targets via interactions with peptidoglycan carbohydrate. We propose that these proteins represent an evolutionarily primitive form of lectin-mediated innate immunity, and that they reveal intestinal strategies for maintaining symbiotic host-microbial relationships. PMID:16931762

  15. Isolation and biochemical characterization of Apios tuber lectin.

    PubMed

    Kenmochi, Eri; Kabir, Syed Rashel; Ogawa, Tomohisa; Naude, Ryno; Tateno, Hiroaki; Hirabayashi, Jun; Muramoto, Koji

    2015-01-09

    Apios tuber lectin, named ATL, was isolated from Apios americana Medikus by two chromatography steps, hydrophobic chromatography and anion-exchange chromatography. The minimum concentration required for the hemagglutination activity toward rabbit erythrocytes of ATL was 4 μg/mL. ATL was composed of a homodimer of 28.4 kDa subunits. The amino acid sequence of ATL was similar to those of other legume lectins. The lectin showed moderate stability toward heating and acidic pH, and the binding affinity against several monosaccharides, such as D-glucosamine and D-galactosamine. ATL also bound to desialylated or agalactosylated glycoproteins such as asialo and agalacto transferrin. ATL decreased the transepithelial electrical resistance across human intestinal Caco-2 cell monolayers, suggesting the effect on the tight junction-mediated paracellular transport.

  16. Parkia pendula lectin as histochemistry marker for meningothelial tumour.

    PubMed

    Beltrão, E I C; Medeiros, P L; Rodrigues, O G; Figueredo-Silva, J; Valença, M M; Coelho, L C B B; Carvalho, L B

    2003-01-01

    Lectins have been intensively used in histochemical techniques for cell surface characterization. These proteins are involved in several biological processes and their use as histochemical markers have been evaluated since they can indicate differences in cell surfaces. Parkia pendula lectin (PpeL) was evaluated as histochemical marker for meningothelial meningioma biopsies. Tissue slices were incubated with PpeL conjugated to horseradish peroxidase (PpeL-HRP) and Concanavalin A-HRP (ConA-HPR) and the binding visualized with diaminobenzidine and hydrogen peroxide. The lectin-tissue binding was inhibited with D-glucose. PpeL showed to be a useful tool for the characterization of meningothelial tumour and clinico-pathological diagnosis.

  17. Purification of a thermostable antinociceptive lectin isolated from Andira anthelmia.

    PubMed

    Nascimento, Kyria Santiago; Nascimento, Francisco Lucas Faustino do; Silva, Mayara Torquato Lima; Nobre, Camila Bezerra; Moreira, Cleane Gomes; Brizeno, Luiz André Cavalcante; da Ponte, Edson Lopes; Assreuy, Ana Maria Sampaio; Cavada, Benildo Sousa

    2016-06-01

    Andira anthelmia (tribe Dalbergieae), a plant from Brazilian Amazon, possesses a seed lectin that was purified by affinity chromatography in sepharose-mannose. This novel Dalbergieae lectin, named AAL, agglutinated rabbit erythrocytes treated with trypsin. The hemagglutinating activity of AAL was maintained after incubation at a wide range of temperature (40 to 70 °C) and pH, was shown to be dependent on divalent cations, and was inhibited by d-mannose and d-sucrose. AAL showed an electrophoretic profile in sodium dodecyl sulfate-polyacrylamide gel electrophoresis similar to other lectins of the tribe Dalbergieae, presenting a double band of molecular weight with approximately 20 kDa and other minor bands of 17, 15, and 13 kDa, being the smaller fragment glycosylated. AAL injected by intravenous route in mice showed antinociceptive activity in two behavioral tests (writhing and formalin). In the writhing test induced by acetic acid, AAL showed inhibitory effect at 0.01 mg/kg (68%), 0.1 mg/kg (46%) and 1 mg/kg (74%). In the formalin test, AAL (0.1 mg/kg) inhibited by 48% the licking time in the inflammatory phase, an effect that was recovered by the lectin association with mannose. In conclusion, AAL presents analgesic effect involving the lectin domain via peripheral mechanisms of inflammatory nociception. This activity highlights the importance of lectins as tools to be used for understanding the interaction of protein-carbohydrate in processes associated to inflammatory pain. Copyright © 2015 John Wiley & Sons, Ltd.

  18. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

    PubMed Central

    Angeli, Renata; da Paz, Nathalia V. N.; Maciel, Jackeline C.; Araújo, Flávia F. B.; Paiva, Patrícia M. G.; Calazans, Glícia M. T.; Valente, Ana Paula; Almeida, Fábio C. L.; Coelho, Luana C. B. B.; Carvalho, Luiz B.; Silva, Maria da Paz C.; dos Santos Correia, Maria Tereza

    2009-01-01

    A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A) and Cratylia mollis (Cramoll 1 and Cramoll 1, 4) did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column. PMID:19547713

  19. Soluble Host Defense Lectins in Innate Immunity to Influenza Virus

    PubMed Central

    Ng, Wy Ching; Tate, Michelle D.; Brooks, Andrew G.; Reading, Patrick C.

    2012-01-01

    Host defenses against viral infections depend on a complex interplay of innate (nonspecific) and adaptive (specific) components. In the early stages of infection, innate mechanisms represent the main line of host defense, acting to limit the spread of virus in host tissues prior to the induction of the adaptive immune response. Serum and lung fluids contain a range of lectins capable of recognizing and destroying influenza A viruses (IAV). Herein, we review the mechanisms by which soluble endogenous lectins mediate anti-IAV activity, including their role in modulating IAV-induced inflammation and disease and their potential as prophylactic and/or therapeutic treatments during severe IAV-induced disease. PMID:22665991

  20. Lectin binding to olfactory system in a shark, Scyliorhinus canicula.

    PubMed

    Franceschini, V; Ciani, F

    1993-01-01

    Lectin histochemical studies were performed on the olfactory system of Scyliorhinus canicula to identify specific glycoconjugates on the cell surface of primary olfactory neurons. The olfactory receptor cells, the olfactory nerve fibers and their terminals in the bulbs were labelled with SBA, BSA-I and BSA-I-B4. The lectin staining patterns indicate that the membranes of small-spotted catshark olfactory neurons had glycoproteins with alpha-galactose residues. This carbohydrate moiety could be related to modulation of the cell-cell interactions in the olfactory system.

  1. Ribosome-inactivating lectins with polynucleotide:adenosine glycosidase activity.

    PubMed

    Battelli, M G; Barbieri, L; Bolognesi, A; Buonamici, L; Valbonesi, P; Polito, L; Van Damme, E J; Peumans, W J; Stirpe, F

    1997-05-26

    Lectins from Aegopodium podagraria (APA), Bryonia dioica (BDA), Galanthus nivalis (GNA), Iris hybrid (IRA) and Sambucus nigra (SNAI), and a new lectin-related protein from Sambucus nigra (SNLRP) were studied to ascertain whether they had the properties of ribosome-inactivating proteins (RIP). IRA and SNLRP inhibited protein synthesis by a cell-free system and, at much higher concentrations, by cells and had polynucleotide:adenosine glycosidase activity, thus behaving like non-toxic type 2 (two chain) RIP. APA and SNAI had much less activity, and BDA and GNA did not inhibit protein synthesis.

  2. Structural and functional similarities of breadfruit seed lectin and jacalin.

    PubMed

    Pineau, N; Pousset, J L; Preud'Homme, J L; Aucouturier, P

    1990-03-01

    Aquous extracts from seeds of Artocarpus altilis (breadfruit) and Artocarpus heterophyllus (jackfruit) were compared by polyacrylamide gel electrophoresis. Two bands of the same size (12 and 15 kD) as the jacalin subunits were the major components in breadfruit seed extract. They strongly reacted with anti-jacalin antibodies by western blotting. The breadfruit lectin displayed the same IgAl and IgD precipitation specificity as jacalin in gel double diffusion experiments. It also stimulated in vitro proliferation of human peripheral blood mononuclear cells. These results suggest that lectins from both species of Artocarpus are very similar.

  3. Interplay between metal binding and cis/trans isomerization in legume lectins: structural and thermodynamic study of P. angolensis lectin.

    PubMed

    Garcia-Pino, Abel; Buts, Lieven; Wyns, Lode; Loris, Remy

    2006-08-04

    The interplay between metal binding, carbohydrate binding activity, stability and structure of the lectin from Pterocarpus angolensis was investigated. Removal of the metals leads to a more flexible form of the protein with significantly less conformational stability. Crystal structures of this metal-free form show significant structural rearrangements, although some structural features that allow the binding of sugars are retained. We propose that substitution of an asparagine residue at the start of the C-terminal beta-strand of the legume lectin monomer hinders the trans-isomerization of the cis-peptide bond upon demetallization and constitutes an intramolecular switch governing the isomer state of the non-proline bond and ultimately the lectin phenotype.

  4. Structures and binding specificity of galactose- and mannose-binding lectins from champedak: differences from jackfruit lectins

    PubMed Central

    Gabrielsen, Mads; Abdul-Rahman, Puteri Shafinaz; Othman, Shatrah; Hashim, Onn H.; Cogdell, Richard J.

    2014-01-01

    Galactose-binding and mannose-binding lectins from the champedak fruit, which is native to South-east Asia, exhibit useful potential clinical applications. The specificity of the two lectins for their respective ligands allows the detection of potential cancer biomarkers and monitoring of the glycosylated state of proteins in human serum and/or urine. To fully understand and expand the use of these natural proteins, their complete sequences and crystal structures are presented here, together with details of sugar binding. PMID:24915077

  5. Lectin-binding sites in epithelial cells of the mouse prostate gland.

    PubMed

    Sakuda, Kentaro; Yoshida, Ayaka; Muragishi, Ryoki; Yoshinaga, Kazuya

    2014-01-01

    The prostate is an exocrine gland in the male reproductive tract that secretes seminal fluids. To gain insight into the cytochemical properties of prostatic epithelial cells, the characteristics of glycoconjugates in mouse prostate sections were examined by lectin histochemistry and immunohistochemistry. Characteristic staining patterns were observed, depending on the type of lectins present in the epithelia. Luminal cells reacted specifically with mannose-binding lectins (Galanthus nivalis lectin, Hippeastrum hybrid lectin, Narcissus pseudonarcissus lectin) and Maclura pomifera lectin in all lobes of the prostate. Luminal cells also expressed galactose, N-acetyl-D-galactosamine (GalNAc), N-acetyl-D-glucosamine (GlcNAc), and fucose residues in the lateral and ventral lobes. Basal cells expressed GlcNAc and fucose, and reacted with Datura stramonium lectin and Aleuria aurantia lectin in all lobes. These results indicate that in the mouse prostate, the selectivity of lectin-binding sites for distinct cell types and lobe-dependent staining may relate to cellular and regional differences in function. Furthermore, some lectins selectively bound to prostatic epithelial cells, indicating their potential use as markers for the histopathological evaluation of prostatic diseases, cancer diagnosis, or male infertility.

  6. Lectin coated MgO nanoparticle: its toxicity, antileishmanial activity, and macrophage activation.

    PubMed

    Jebali, Ali; Hekmatimoghaddam, Seyedhossein; Kazemi, Bahram; Allaveisie, Azra; Masoudi, Alireza; Daliri, Karim; Sedighi, Najme; Ranjbari, Javad

    2014-10-01

    The purpose of this research was to evaluate toxicity of uncoated magnesium oxide nanoparticles (MgO NPs), MgO NPs coated with Peanut agglutinin (PNA) lectin, and PNA alone on the promastigotes of Leishmania major (L. major) and macrophages of BALB/c mice. On the other hand, antileishmanial property of uncoated MgO NPs, lectin coated MgO NPs, and PNA lectin alone was evaluated, and also macrophage activation was investigated after treatment with these materials by measurement of nitrite, H2O2, and some interleukins. This study showed that PNA lectin and lectin coated MgO NPs had approximately no toxicity on L. major and macrophages, but some toxic effects were observed for uncoated MgO NPs, especially at concentration of 500 µg/mL. Interestingly, lectin coated MgO NPs had the highest antileishmanial activity and macrophage activation, compared with uncoated MgO NPs and PNA lectin.

  7. The use of lectin microarray for assessing glycosylation of therapeutic proteins

    PubMed Central

    Zhang, Lei; Luo, Shen; Zhang, Baolin

    2016-01-01

    ABSTRACT Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. However, the complexity of protein glycosylation poses a daunting analytical challenge. In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. PMID:26918373

  8. The use of lectin microarray for assessing glycosylation of therapeutic proteins.

    PubMed

    Zhang, Lei; Luo, Shen; Zhang, Baolin

    2016-01-01

    Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. However, the complexity of protein glycosylation poses a daunting analytical challenge. In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins.

  9. Purification and characterization of a new type lactose-binding Ulex europaeus lectin by affinity chromatography.

    PubMed

    Konami, Y; Yamamoto, K; Osawa, T

    1991-02-01

    A new type lactose-binding lectin was purified from extracts of Ulex europaeus seeds by affinity chromatography on a column of galactose-Sepharose 4B, followed by gel filtration on Sephacryl S-300. This lectin, designated as Ulex europaeus lectin III (UEA-III), was found to be inhibited by lactose. The dimeric lectin is a glycoprotein with a molecular mass of 70,000 Da; it consists of two apparently identical subunits of a molecular mass of 34,000 Da. Compositional analysis showed that this lectin contains 30% carbohydrate and a large amount of aspartic acid, serine and valine, but no sulfur-containing amino acids. The N-terminal amino-acid sequences of L-fucose-binding Ulex europaeus lectin I (UEA-I) and di-N-acetylchitobiose-binding Ulex europaeus lectin II (UEA-II), both of which we have already purified and characterized, and that of UEA-III were determined and compared.

  10. Use of lectin microarray to differentiate gastric cancer from gastric ulcer

    PubMed Central

    Huang, Wei-Li; Li, Yang-Guang; Lv, Yong-Chen; Guan, Xiao-Hui; Ji, Hui-Fan; Chi, Bao-Rong

    2014-01-01

    AIM: To investigate the feasibility of lectin microarray for differentiating gastric cancer from gastric ulcer. METHODS: Twenty cases of human gastric cancer tissue and 20 cases of human gastric ulcer tissue were collected and processed. Protein was extracted from the frozen tissues and stored. The lectins were dissolved in buffer, and the sugar-binding specificities of lectins and the layout of the lectin microarray were summarized. The median of the effective data points for each lectin was globally normalized to the sum of medians of all effective data points for each lectin in one block. Formalin-fixed paraffin-embedded gastric cancer tissues and their corresponding gastric ulcer tissues were subjected to Ag retrieval. Biotinylated lectin was used as the primary antibody and HRP-streptavidin as the secondary antibody. The glycopatterns of glycoprotein in gastric cancer and gastric ulcer specimens were determined by lectin microarray, and then validated by lectin histochemistry. Data are presented as mean ± SD for the indicated number of independent experiments. RESULTS: The glycosylation level of gastric cancer was significantly higher than that in ulcer. In gastric cancer, most of the lectin binders showed positive signals and the intensity of the signals was stronger, whereas the opposite was the case for ulcers. Significant differences in the pathological score of the two lectins were apparent between ulcer and gastric cancer tissues using the same lectin. For MPL and VVA, all types of gastric cancer detected showed stronger staining and a higher positive rate in comparison with ulcer, especially in the case of signet ring cell carcinoma and intra-mucosal carcinoma. GalNAc bound to MPL showed a significant increase. A statistically significant association between MPL and gastric cancer was observed. As with MPL, there were significant differences in VVA staining between gastric cancer and ulcer. CONCLUSION: Lectin microarray can differentiate the different

  11. Use of lectin microarray to differentiate gastric cancer from gastric ulcer.

    PubMed

    Huang, Wei-Li; Li, Yang-Guang; Lv, Yong-Chen; Guan, Xiao-Hui; Ji, Hui-Fan; Chi, Bao-Rong

    2014-05-14

    To investigate the feasibility of lectin microarray for differentiating gastric cancer from gastric ulcer. Twenty cases of human gastric cancer tissue and 20 cases of human gastric ulcer tissue were collected and processed. Protein was extracted from the frozen tissues and stored. The lectins were dissolved in buffer, and the sugar-binding specificities of lectins and the layout of the lectin microarray were summarized. The median of the effective data points for each lectin was globally normalized to the sum of medians of all effective data points for each lectin in one block. Formalin-fixed paraffin-embedded gastric cancer tissues and their corresponding gastric ulcer tissues were subjected to Ag retrieval. Biotinylated lectin was used as the primary antibody and HRP-streptavidin as the secondary antibody. The glycopatterns of glycoprotein in gastric cancer and gastric ulcer specimens were determined by lectin microarray, and then validated by lectin histochemistry. Data are presented as mean ± SD for the indicated number of independent experiments. The glycosylation level of gastric cancer was significantly higher than that in ulcer. In gastric cancer, most of the lectin binders showed positive signals and the intensity of the signals was stronger, whereas the opposite was the case for ulcers. Significant differences in the pathological score of the two lectins were apparent between ulcer and gastric cancer tissues using the same lectin. For MPL and VVA, all types of gastric cancer detected showed stronger staining and a higher positive rate in comparison with ulcer, especially in the case of signet ring cell carcinoma and intra-mucosal carcinoma. GalNAc bound to MPL showed a significant increase. A statistically significant association between MPL and gastric cancer was observed. As with MPL, there were significant differences in VVA staining between gastric cancer and ulcer. Lectin microarray can differentiate the different glycopatterns in gastric cancer and

  12. Stimulation of vascular cell proliferation by beta-galactoside specific lectins.

    PubMed

    Sanford, G L; Harris-Hooker, S

    1990-08-01

    An investigation was conducted to assess the effects of various beta-galactoside specific lectins on the growth of vascular cells in vitro. The plant lectins from peanut (Arachis hypogaea), mushroom (Agaricus bisporus), and coral tree (Erythrina corallodendron) were used in these studies with the ultimate purpose of comparing those findings with data derived with the lectin isolated from rat lung. Peanut lectin was added to confluent and subconfluent cultures of smooth muscle cells (SMC), pulmonary arterial (PEC), and aortic endothelial cells (BAEC) at concentrations of 2, 3.5, and 7.0 micrograms/ml. There was a dose-dependent increase in cell proliferation for both confluent and subconfluent SMC, with maximal stimulation noted between 3.5 and 7 micrograms/ml of peanut lectin. A dose-dependent stimulation of PEC proliferation was also found with maximal stimulation between 3.5 and 7.0 micrograms/ml. Peanut lectin did not stimulate BAEC to multiply. The stimulation of PEC and SMC by peanut lectin could be prevented by the addition of 50 mM lactose. Peanut and mushroom lectin stimulated the proliferation of sparse cultures of SMC in a dose-dependent fashion in both standard (10% fetal bovine serum, or FBS) or low (0.5% FBS) serum to about the same degree. Coral tree lectin did not have a significant stimulation of proliferation under either serum conditions. The incorporation of [3H]thymidine into the DNA of PEC was increased 30 and 150% by peanut lectin and lung galaptin, respectively, under standard serum conditions. However, under low serum conditions, both lectins increased incorporation by about the same extent (93 and 78% for peanut lectin and galaptin, respectively). Both lectins produced a 30% increase in DNA synthesis by SMC under standard serum conditions, and about a 200% increase under low serum conditions. These studies indicate that beta-galactoside specific lectins such as lung galaptin have mitogenic activity toward vascular cells.

  13. Molecular basis for peptidoglycan recognition by a bactericidal lectin.

    PubMed

    Lehotzky, Rebecca E; Partch, Carrie L; Mukherjee, Sohini; Cash, Heather L; Goldman, William E; Gardner, Kevin H; Hooper, Lora V

    2010-04-27

    RegIII proteins are secreted C-type lectins that kill Gram-positive bacteria and play a vital role in antimicrobial protection of the mammalian gut. RegIII proteins bind their bacterial targets via interactions with cell wall peptidoglycan but lack the canonical sequences that support calcium-dependent carbohydrate binding in other C-type lectins. Here, we use NMR spectroscopy to determine the molecular basis for peptidoglycan recognition by HIP/PAP, a human RegIII lectin. We show that HIP/PAP recognizes the peptidoglycan carbohydrate backbone in a calcium-independent manner via a conserved "EPN" motif that is critical for bacterial killing. While EPN sequences govern calcium-dependent carbohydrate recognition in other C-type lectins, the unusual location and calcium-independent functionality of the HIP/PAP EPN motif suggest that this sequence is a versatile functional module that can support both calcium-dependent and calcium-independent carbohydrate binding. Further, we show HIP/PAP binding affinity for carbohydrate ligands depends on carbohydrate chain length, supporting a binding model in which HIP/PAP molecules "bind and jump" along the extended polysaccharide chains of peptidoglycan, reducing dissociation rates and increasing binding affinity. We propose that dynamic recognition of highly clustered carbohydrate epitopes in native peptidoglycan is an essential mechanism governing high-affinity interactions between HIP/PAP and the bacterial cell wall.

  14. Molecular basis for peptidoglycan recognition by a bactericidal lectin

    PubMed Central

    Lehotzky, Rebecca E.; Partch, Carrie L.; Mukherjee, Sohini; Cash, Heather L.; Goldman, William E.; Gardner, Kevin H.; Hooper, Lora V.

    2010-01-01

    RegIII proteins are secreted C-type lectins that kill Gram-positive bacteria and play a vital role in antimicrobial protection of the mammalian gut. RegIII proteins bind their bacterial targets via interactions with cell wall peptidoglycan but lack the canonical sequences that support calcium-dependent carbohydrate binding in other C-type lectins. Here, we use NMR spectroscopy to determine the molecular basis for peptidoglycan recognition by HIP/PAP, a human RegIII lectin. We show that HIP/PAP recognizes the peptidoglycan carbohydrate backbone in a calcium-independent manner via a conserved “EPN” motif that is critical for bacterial killing. While EPN sequences govern calcium-dependent carbohydrate recognition in other C-type lectins, the unusual location and calcium-independent functionality of the HIP/PAP EPN motif suggest that this sequence is a versatile functional module that can support both calcium-dependent and calcium-independent carbohydrate binding. Further, we show HIP/PAP binding affinity for carbohydrate ligands depends on carbohydrate chain length, supporting a binding model in which HIP/PAP molecules “bind and jump” along the extended polysaccharide chains of peptidoglycan, reducing dissociation rates and increasing binding affinity. We propose that dynamic recognition of highly clustered carbohydrate epitopes in native peptidoglycan is an essential mechanism governing high-affinity interactions between HIP/PAP and the bacterial cell wall. PMID:20382864

  15. Architectures of Multivalent Glycomimetics for Probing Carbohydrate-Lectin Interactions

    NASA Astrophysics Data System (ADS)

    Lahmann, Martina

    Well-defined multivalent glycoconjugates are valued tools in glycoscience and they are particularly valuable for the investigation of carbohydrate-lectin interactions. In addition to the relatively globularly shaped glycodendrimers many other designs have been realized. This chapter gives an overview on the common different architectures and their chemical synthesis by focussing on the achievements made since 2001.

  16. Lectins: a primer for histochemists and cell biologists.

    PubMed

    Manning, Joachim C; Romero, Antonio; Habermann, Felix A; García Caballero, Gabriel; Kaltner, Herbert; Gabius, Hans-Joachim

    2017-02-01

    An experimental observation on selecting binding partners underlies the introduction of the term 'lectin'. Agglutination of erythrocytes depending on their blood-group status revealed the presence of activities in plant extracts that act in an epitope-specific manner like antibodies. As it turned out, their binding partners on the cell surface are carbohydrates of glycoconjugates. By definition, lectins are glycan-specific (mono- or oligosaccharides presented by glycoconjugates or polysaccharides) receptors, distinguished from antibodies, from enzymes using carbohydrates as substrates and from transporters of free saccharides. They are ubiquitous in Nature and structurally widely diversified. More than a dozen types of folding pattern have evolved for proteins that bind glycans. Used as tool, this capacity facilitates versatile mapping of glycan presence so that plant/fungal and also animal/human lectins have found a broad spectrum of biomedical applications. The functional pairing with physiological counterreceptors is involved in a wide range of cellular activities from cell adhesion, glycoconjugate trafficking to growth regulation and lets lectins act as sensors/effectors in host defense.

  17. Pseudomonas Aeruginosa Lectins As Targets for Novel Antibacterials

    PubMed Central

    Grishin, A. V.; Krivozubov, M. S.; Karyagina, A. S.; Gintsburg, A. L.

    2015-01-01

    Pseudomonas aeruginosa is one of the most widespread and troublesome opportunistic pathogens that is capable of colonizing various human tissues and organs and is often resistant to many currently used antibiotics. This resistance is caused by different factors, including the acquisition of specific resistance genes, intrinsic capability to diminish antibiotic penetration into the bacterial cell, and the ability to form biofilms. This situation has prompted the development of novel compounds differing in their mechanism of action from traditional antibiotics that suppress the growth of microorganisms or directly kill bacteria. Instead, these new compounds should decrease the pathogens’ ability to colonize and damage human tissues by inhibiting the virulence factors and biofilm formation. The lectins LecA and LecB that bind galactose and fucose, as well as oligo- and polysaccharides containing these sugars, are among the most thoroughly-studied targets for such novel antibacterials. In this review, we summarize the results of experiments highlighting the importance of these proteins for P. aeruginosa pathogenicity and provide information on existing lectins inhibitors and their effectiveness in various experimental models. Particular attention is paid to the effects of lectins inhibition in animal models of infection and in clinical practice. We argue that lectins inhibition is a perspective approach to combating P. aeruginosa. However, despite the existence of highly effective in vitro inhibitors, further experiments are required in order to advance these inhibitors into pre-clinical studies. PMID:26085942

  18. Momordica charantia seed lectin: toxicity, bacterial agglutination and antitumor properties.

    PubMed

    Kabir, Syed Rashel; Nabi, Md Mahamodun; Nurujjaman, Md; Abu Reza, Md; Alam, A H M Khurshid; Uz Zaman, Rokon; Khalid-Bin-Ferdaus, Khandaker Md; Amin, Ruhul; Khan, Md Masudul Hasan; Hossain, Md Anowar; Uddin, Md Salim; Mahmud, Zahid Hayat

    2015-03-01

    In last three decades, several studies were carried out on the D-galactose-specific lectin of Momordica charantia seeds (MCL). In the present study, in vitro growth inhibition (8-23 %) at different concentrations (6-24 μg/ml) of MCL was observed against Ehrlich ascites carcinoma (EAC) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MCL also showed 28, 45, and 75 % growth inhibitions against EAC cells when administered 1.2, 2.0, and 2.8 mg/kg/day (i.p.), respectively for five consequent days in vivo in mice. After lectin treatment, the level of red blood cell and hemoglobin was increased significantly with the decrease of white blood cell and maintained the normal level when compared with EAC-bearing control and normal mice without EAC cells. Although MCL caused cell cycle arrest at G0/G1 phase of EAC cells, any irregular shape or apoptotic morphological alterations in the lectin-treated EAC cells was not observed by an optical and fluorescence microscope. Lectin showed toxicity against brine shrimp nauplii with an LC50 value of 49.7 μg/ml. Four out of seven pathogenic bacteria were agglutinated by MCL in the absence of inhibitory sugar D-lactose/D-galactose. In conclusion, MCL showed strong cytotoxic effect and therefore can be used as a potent anticancer chemotherapeutic agent.

  19. Lectin Binding to Radopholus citrophilus and R. similis Proteins.

    PubMed

    Kaplan, D T; Gottwald, T R

    1992-06-01

    Lectin-binding glycoproteins in seven populations of two burrowing nematode sibling species were probed with five different biotinylated lectins on Western blots, and differences were correlated with nematode ability to parasitize citrus and to overcome citrus rootstock resistance. Banding patterns of molecular weight standards were fit best by an exponential decay function, and a predictive equation was used to estimate molecular weights (r(2) = 0.999). A band (131 kDa) that labeled with the lectin Concanavalin A (Con A) occurred in extracts from cuticles and egg shells of populations of Radopholus citrophilus that parasitize citrus. Wheat germ agglutin labeled a band (58 kDa) in aqueous homogenates of populations that reproduce in roots of citrus rootstock normally resistant to burrowing nematodes. The two sibling species R. citrophilus and R. similis were distinguished by a high molecular weight Con A-labeled band (608 kDa) from cuticle and egg shells. Probing blots with the lectin Limulus polyphemus agglutinin indicated that each population contained a band (12-16 kDa) specifically inhibited by the addition of 25 mM neuraminic acid, suggesting that glycoproteins with sialic acid moieties are present in burrowing nematodes.

  20. [Purification of IgY against Salvia bogotensis lectin].

    PubMed

    Barroso, Paola; Murcia, Hansen; Vega, Nohora; Pérez, Gerardo

    2005-12-01

    Egg yolk immunoglobulins (IgY) have been extensively used in immunology, biochemistry, and biotechnology in studies of human and animal health. However, their use requires two preparatory steps: first, the egg yolk lipids must be removed without impairing the immunoglobin functional properties, and second, the isolation methods must allow high recoveries for further use as detection and purification tools. Because Tn antigen presence serves as a tumoral marker and because S. bogotensis lectin's can specifically recognize this antigen, the current study aims at making available an anti-lectin IgY. This tool will be useful in histochemical and cellular studies involving transformed cells. Anti-lectin IgY was produced by immunization of hens with S. bogotensis lectin, and the effect of antigen dose on IgY levels was assesed. Several methods for lipid removal, IgY extraction and purification were assayed, and yields and purity of IgYs were established for each method. The best delipidation and extraction method included yolk dilution with water under acidic conditions and (NH4)2SO4 60% s precipitation from which 43 mg protein/yolk were recovered. Among the chromatographic methods, thiophilic chromatography permitted the recovery of a substantial quantity of pure IgY (10.4 mg IgY/yolk). With this method, the function and characteristics of IgY were preserved. The best conditions for anti-S. bogotensis functional IgY extraction and purification were established.

  1. Genetics Home Reference: mannose-binding lectin deficiency

    MedlinePlus

    ... MBL2 gene. Mannose-binding lectin plays an important role in the body's immune response by attaching to foreign invaders such as bacteria, viruses, or yeast and turning on (activating) the complement system . The complement system is a group of immune system proteins that work together to ...

  2. Peanut lectin-binding sites in large bowel carcinoma.

    PubMed

    Cooper, H S

    1982-10-01

    Peanut lectin is known to bind to B-D-Gal-(1 leads to 3)-D-GalNac which provides antigenic determination for the T (TAg) blood group antigen. We examined 33 rectosigmoid carcinomas and 15 corresponding controls for their ability to express peanut lectin-binding sites. In controls one could localize TAg to the supranuclear portion of the cell, however, in cancers one noticed a cytostructural relocalization of TAg with the following two major patterns: localization to the region of the glycocalyx and localization intracytoplasmically in the apical portion of the cell. These two patterns were associated with glandular differentiation. Less frequently noted or in association with the above was a mucin glob-like pattern and/or a fine diffuse intracytoplasmic pattern associated with solid, nonglandular areas. The more poorly differentiated cancers less frequently expressed peanut lectin-binding sites. Benign (nontransitional zone) epithelium in those patients whose tumor expressed TAg was negative for peanut lectin-binding sites in 66 per cent of the cases. Reduced tumoral glycosyltransferases may explain this increased synthesis of TAg in cancers as compared with controls, if one considers TAg to be an incomplete glycoprotein of the MN blood group system.

  3. Crystallization and preliminary characterization of a highly thermostable lectin from Trichosanthes dioica and comparison with other Trichosanthes lectins

    SciTech Connect

    Dharkar, Poorva D.; Anuradha, P.; Gaikwad, Sushama M.; Suresh, C. G.

    2006-03-01

    A lectin from Trichosanthes dioica seeds has been purified and crystallized using 25%(w/v) PEG 2K MME, 0.2 M ammonium acetate, 0.1 M Tris–HCl pH 8.5 and 50 µl 0.5%(w/v) n-octyl β-d-glucopyranoside as thick needles belonging to hexagonal space group P6{sub 4}. A lectin from Trichosanthes dioica seeds has been purified and crystallized using 25%(w/v) PEG 2K MME, 0.2 M ammonium acetate, 0.1 M Tris–HCl pH 8.5 and 50 µl 0.5%(w/v) n-octyl β-d-glucopyranoside as thick needles belonging to hexagonal space group P6{sub 4}. Unit-cell parameters were a = b = 167.54, c = 77.42 Å. The crystals diffracted to a Bragg spacing of 2.8 Å. Both the structures of abrin-a and T. kirilowii lectin could be used as a model in structure determination using the molecular-replacement method; however, T. kirilowii lectin coordinates gave better values of reliability and correlation parameters. The thermal, chemical and pH stability of this lectin have also been studied. When heated, its haemagglutination activity remained unaffected up to 363 K. Other stability studies show that 4 M guanidinium hydrochloride (Gdn–HCl) initiates unfolding and that the protein is completely unfolded at 6 M Gdn–HCl. Treatment with urea resulted in a total loss of activity at higher concentrations of denaturant with no major structural changes. The protein remained stable over a wide pH range, from pH 6 to pH 12, except for partial unfolding at extremely alkaline pH. The role of disulfide bonds in the protein stability was found to be insignificant. Rayleigh light-scattering studies showed no molecular aggregation in any of the extreme treated conditions. The unusual stability of this lectin resembles that of type II ribosome-inactivating proteins (type II RIPs), which is also supported by structure determination. The structural features observed in a preliminary electron-density map were compared with the other two available Trichosanthes lectin structures.

  4. Visualizing the dental biofilm matrix by means of fluorescence lectin-binding analysis

    PubMed Central

    Tawakoli, Pune N.; Neu, Thomas R.; Busck, Mette M.; Kuhlicke, Ute; Schramm, Andreas; Attin, Thomas; Wiedemeier, Daniel B.; Schlafer, Sebastian

    2017-01-01

    ABSTRACT The extracellular matrix is a poorly studied, yet important component of dental biofilms. Fluorescence lectin-binding analysis (FLBA) is a powerful tool to characterize glycoconjugates in the biofilm matrix. This study aimed to systematically investigate the ability of 75 fluorescently labeled lectins to visualize and quantify extracellular glycoconjugates in dental biofilms. Lectin binding was screened on pooled supragingival biofilm samples collected from 76 subjects using confocal microscopy. FLBA was then performed with 10 selected lectins on biofilms grown in situ for 48 h in the absence of sucrose. For five lectins that proved particularly suitable, stained biovolumes were quantified and correlated to the bacterial composition of the biofilms. Additionally, combinations of up to three differently labeled lectins were tested. Of the 10 lectins, five bound particularly well in 48-h-biofilms: Aleuria aurantia (AAL), Calystega sepiem (Calsepa), Lycopersicon esculentum (LEA), Morniga-G (MNA-G) and Helix pomatia (HPA). No significant correlation between the binding of specific lectins and bacterial composition was found. Fluorescently labeled lectins enable the visualization of glycoconjugates in the dental biofilm matrix. The characterization and quantification of glycoconjugates in dental biofilms require a combination of several lectins. For 48-h-biofilms grown in absence of sucrose, AAL, Calsepa, HPA, LEA, and MNA-G are recommendable. PMID:28748044

  5. Isolation and partial characterization of a lectin from ground elder (Aegopodium podagraria) rhizomes.

    PubMed

    Peumans, W J; Nsimba-Lubaki, M; Peeters, B; Broekaert, W F

    1985-05-01

    A lectin has been isolated from rhizomes of ground elder (Aegopodium podagraria) using a combination of affinity chromatography on erythrocyte membrane proteins immobilized on cross-linked agarose and hydroxyapatite, and ion-exchange chromatography. The molecular structure of the lectin was determined by gelfiltration, sucrose density-gradient centrifugation and gel electrophoresis under denaturing conditions. It has an unusually high Mr (about 480000) and is most probably an octamer composed of two distinct types of subunits with slightly different Mr (about 60000). Hapten inhibition assays indicated that the Aegopodium lectin is preferentially inhibited by N-acetylgalactosamine. Nevertheless, it does not agglutinate preferentially blood-group-A erythrocytes. The ground-elder lectin is a typical non-seed lectin, which occurs virtually exclusively in the underground rhizomes. In this organ it is an abundant protein as it represents up to 5% of the total protein content. The lectin content of the rhizome tissue varies strongly according to its particular location along the organ. In addition, the lectin content changes dramatically as a function of the seasons. The ground-elder lectin differs from all other plant lectins by its unusually high molecular weight. In addition, it is the first lectin to be isolated from a species of the family Apiaceae.

  6. Interactions of lectins with plasma membrane glycoproteins of the Ehrlich ascites carcinoma cell.

    PubMed

    Nachbar, M S; Oppenheim, J D; Aull, F

    1976-02-06

    Several aspects of the interaction of various lectins with the surface of Ehrlich ascites carcinoma cells are described. The order of agglutinating activity for various lectins is Ricinus communis greater than wheat germ greater than or equal to concanavalin A greater than or equal to soybean greater than Limulus polyphemus. No agglutination was noted for Ulex europaeus. Using 125I-labeled lectins it was determined that there are 1.6 and 7 times as many Ricinus communis lectin binding sites for concanavalin A and soybean lectins. Sodium deoxycholate-solubilized plasma membrane material was subjected to lectin affinity chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lectin receptors of the plasma membrane appeared to be heterogeneous and some qualitative differences could be discerned among the electrophoretically analyzed material, which bound to and was specifically eluted from the various lectin affinity columns. The characteristics of elution of bound material from individual lectin columns indicated secondary hydrophobic interactions between concanavalin A or wheat germ agglutinin and their respective lectin receptor molecules.

  7. Selective binding of lectins to normal and neoplastic urothelium in rat and mouse bladder carcinogenesis models.

    PubMed

    Zupančič, Daša; Kreft, Mateja Erdani; Romih, Rok

    2014-01-01

    Bladder cancer adjuvant intravesical therapy could be optimized by more selective targeting of neoplastic tissue via specific binding of lectins to plasma membrane carbohydrates. Our aim was to establish rat and mouse models of bladder carcinogenesis to investigate in vivo and ex vivo binding of selected lectins to the luminal surface of normal and neoplastic urothelium. Male rats and mice were treated with 0.05 % N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in drinking water and used for ex vivo and in vivo lectin binding experiments. Urinary bladder samples were also used for paraffin embedding, scanning electron microscopy and immunofluorescence labelling of uroplakins. During carcinogenesis, the structure of the urinary bladder luminal surface changed from microridges to microvilli and ropy ridges and the expression of urothelial-specific glycoproteins uroplakins was decreased. Ex vivo and in vivo lectin binding experiments gave comparable results. Jacalin (lectin from Artocarpus integrifolia) exhibited the highest selectivity for neoplastic compared to normal urothelium of rats and mice. The binding of lectin from Amaranthus caudatus decreased in rat model and increased in mouse carcinogenesis model, indicating interspecies variations of plasma membrane glycosylation. Lectin from Datura stramonium showed higher affinity for neoplastic urothelium compared to the normal in rat and mouse model. The BBN-induced animal models of bladder carcinogenesis offer a promising approach for lectin binding experiments and further lectin-mediated targeted drug delivery research. Moreover, in vivo lectin binding experiments are comparable to ex vivo experiments, which should be considered when planning and optimizing future research.

  8. Properties of Lectins in the Root and Seed of Lotononis bainesii1

    PubMed Central

    Law, Ian J.; Strijdom, Barend W.

    1984-01-01

    A lectin was purified from the root of Lotononis bainesii Baker by affinity chromatography on Sepharose-blood group substance A + H. The molecular weight of the lectin was estimated by gel filtration to be 118,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the lectin was a tetramer composed of two slightly different subunits with respective molecular weights of 32,000 and 35,000. The lectin had a hexose content of 12% (w/w) and contained the sugars fucose, glucosamine, mannose, and xylose. Root lectin hemagglutination was preferentially inhibited by disaccharides with terminal nonreducing galactose residues. Antigens capable of cross-reaction with root lectin antibody were not detected in the seed of L. bainesii. A lectin from the seed of L. bainesii was partially purified by adsorption to pronase-treated rabbit erythrocytes. The lectin preparation had a molecular weight of approximately 200,000. Galactose and galactono-1,4-lactone inhibited seed lectin hemagglutination but lactose was ineffective. There was no evidence that the root of L. bainesii contained material antigenically related to the seed lectin. Images Fig. 2 Fig. 4 Fig. 5 PMID:16663508

  9. Microbial F-type lectin domains with affinity for blood group antigens.

    PubMed

    Mahajan, Sonal; Khairnar, Aasawari; Bishnoi, Ritika; Ramya, T N C

    2017-09-23

    F-type lectins are fucose binding lectins with characteristic fucose binding and calcium binding motifs. Although they occur with a selective distribution in viruses, prokaryotes and eukaryotes, most biochemical studies have focused on vertebrate F-type lectins. Recently, using sensitive bioinformatics search techniques on the non-redundant database, we had identified many microbial F-type lectin domains with diverse domain organizations. We report here the biochemical characterization of F-type lectin domains from Cyanobium sp. PCC 7001, Myxococcus hansupus and Leucothrix mucor. We demonstrate that while all these three microbial F-type lectin domains bind to the blood group H antigen epitope on fucosylated glycans, there are fine differences in their glycan binding specificity. Cyanobium sp. PCC 7001 F-type lectin domain binds exclusively to extended H type-2 motif, Myxococcus hansupus F-type lectin domain binds to B, H type-1 and Lewis(b) motifs, and Leucothrix mucor F-type lectin domain binds to a wide range of fucosylated glycans, including A, B, H and Lewis antigens. We believe that these microbial lectins will be useful additions to the glycobiologist's toolbox for labeling, isolating and visualizing glycans. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Disruption of the C. elegans Intestinal Brush Border by the Fungal Lectin CCL2 Phenocopies Dietary Lectin Toxicity in Mammals

    PubMed Central

    Stutz, Katrin; Kaech, Andres; Aebi, Markus; Künzler, Markus; Hengartner, Michael O.

    2015-01-01

    Lectins are non-immunoglobulin carbohydrate-binding proteins without enzymatic activity towards the bound carbohydrates. Many lectins of e.g. plants or fungi have been suggested to act as toxins to defend the host against predators and parasites. We have previously shown that the Coprinopsis cinerea lectin 2 (CCL2), which binds to α1,3-fucosylated N-glycan cores, is toxic to Caenorhabditis elegans and results in developmental delay and premature death. In this study, we investigated the underlying toxicity phenotype at the cellular level by electron and confocal microscopy. We found that CCL2 directly binds to the intestinal apical surface and leads to a highly damaged brush border with loss of microvilli, actin filament depolymerization, and invaginations of the intestinal apical plasma membrane through gaps in the terminal web. We excluded several possible toxicity mechanisms such as internalization and pore-formation, suggesting that CCL2 acts directly on intestinal apical plasma membrane or glycocalyx proteins. A genetic screen for C. elegans mutants resistant to CCL2 generated over a dozen new alleles in bre 1, ger 1, and fut 1, three genes required for the synthesis of the sugar moiety recognized by CCL2. CCL2-induced intestinal brush border defects in C. elegans are similar to the damage observed previously in rats after feeding the dietary lectins wheat germ agglutinin or concanavalin A. The evolutionary conserved reaction of the brush border between mammals and nematodes might allow C. elegans to be exploited as model organism for the study of dietary lectin-induced intestinal pathology in mammals. PMID:26057124

  11. Purification of the glycoprotein lectin from the broad bean (Vicia faba) and a comparison of its properties with lectins of similar specificity.

    PubMed Central

    Allen, A K; Desai, N N; Neuberger, A

    1976-01-01

    1. The lectin from the broad bean (Vicia faba) was purified by affinity chromatography by using 3-O-methylglucosamine covalently attached through the amino group to CH-Sepharose (an omega-hexanoic acid derivative of agarose). Its composition and the nature of its subunits were compared with concanavalin A and the lectins from pea and lentil. 2. Unlike the other three lectins, broad-bean lectin is a glycoprotein; a glycopeptide containing glucosamine and mannose was isolated from a proteolytic digest. 3. The mol.wt. is about 47500; the glycoprotein consists of two apprently identical subunits, held together by non-covalent forces. Fragments of the subunits, similar to those found in concanavalin A and soya-bean agglutinin, were found in active preparations. 4. Broad-bean lectin was compared with concanavalin A and the lectins from pea and lentil in an investigation of the inhibition of their action by a number of monosaccharides, methyl ethers of monosaccharides, disaccharides and glycopeptides. The most striking differences concern 3-O-substituted monosaccharides, which are strong inhibitors of the action of broad-bean, pea and lentil lectins but not of the action of concanavalin A. There is, however, no strong inhibition of the action of these lectins by 3-Olinked disaccharides. PMID:938471

  12. Blocking of Pseudomonas aeruginosa and Chromobacterium violaceum lectins by diverse mammalian milks.

    PubMed

    Zinger-Yosovich, K D; Iluz, D; Sudakevitz, D; Gilboa-Garber, N

    2010-02-01

    Pseudomonas aeruginosa and Chromobacterium violaceum morbid and mortal infections are initiated by bacterial adherence to host-cell receptors via their adhesins, including lectins (which also contribute to bacterial biofilm formation). Pseudomonas aeruginosa produces a galactophilic lectin, PA-IL (LecA), and a fucophilic (Lewis-specific) lectin, PA-IIL (LecB), and C. violaceum produces a fucophilic (H-specific) lectin, CV-IIL. The antibiotic resistance of these bacteria prompted the search for glycosylated receptor-mimicking compounds that would function as glycodecoys for blocking lectin attachment to human cell receptors. Lectins PA-IL and PA-IIL have been shown to be useful for such glycodecoy probing, clearly differentiating between human and cow milks. This article describes their usage, together with CV-IIL and the plant lectin concanavalin A, for comparing the anti-lectin-dependent adhesion potential of diverse mammalian milks. The results show that the diverse milks differ in blocking (hemagglutination inhibition) and differential binding (Western blots) of these lectins. Human milk most strongly inhibited the 3 bacterial lectins (with PA-IIL superiority), followed by alpaca, giraffe, and monkey milks, whereas cow milk was a weak inhibitor. Lectin PA-IL was inhibited strongly by human, followed by alpaca, mare, giraffe, buffalo, and monkey milks, weakly by camel milk, and not at all by rabbit milk. Lectins PA-IIL and CV-IIL were also most sensitive to human milk, followed by alpaca, monkey, giraffe, rabbit, and camel milks but negligibly sensitive to buffalo and mare milks. Plant lectin concanavalinA, which was used as the reference, differed from them in that it was much less sensitive to human milk and was equally as sensitive to cow milk. These results have provided important information on the anti-lectin-dependent adhesion potential of the diverse milks examined. They showed that human followed by alpaca, giraffe, and Rhesus monkey milks efficiently

  13. [Extracellular lectins from saprophytic strains of bacteria of the genus Bacillus (review)].

    PubMed

    Podgorskiĭ, V S; Kovalenko, É A; Karpova, I S; Sashchuk, E V; Get'man, E I

    2014-01-01

    The present review summarizes both the authors' own and other researchers' reports concerning the synthesis and properties of sialic acid-specific extracellular lectins from saprophytic bacteria of the genus Bacillus. Bacilli of this genus isolated from different ecological niches differ with regard to the ability to synthesize lectins. The biphasic temporal pattern of lectin synthesis in culture and the effect of cultivation conditions on lectin production have been demonstrated. The advantages of the technological procedure used for the isolation and purification of these biopolymers have been characterized. Specificity towards sialic acid, which is characteristic only of a small number of lectins from bacilli and underlies a wide range of biological effects of these proteins, receives particular attention. Adaptation mechanisms involving lectins--carbohydrate-recognizing proteins--are suggested to have developed in saprophytic bacilli to ensure the survival of these organisms in a constantly changing environment.

  14. Purification, crystallization and preliminary X-ray crystallographic analysis of rice lectin from Oryza sativa

    SciTech Connect

    Huang, Yen-Chieh; Lin, Yi-Hung; Shih, Chia-Hao; Shih, Chun-Liang; Chang, Tschining; Chen, Chun-Jung

    2006-02-01

    Rice lectin was crystallized and analyzed by X-ray crystallography. Lectins with sugar-binding specificity are widely distributed in higher plants and various other species. The expression of rice lectin from Oryza sativa is up-regulated in the growing coleoptile when anaerobic stress persists. A rice lectin of molecular weight 15.2 kDa has been crystallized using the hanging-drop vapour-diffusion method. From the diffraction of the lectin crystals at 1.93 Å resolution, the unit cell belongs to space group P3{sub 1}, with unit-cell parameters a = 98.58, b = 98.58, c = 44.72 Å. Preliminary analysis indicates that there are two lectin molecules in an asymmetric unit with a large solvent content, 70.1%.

  15. [Lectin-binding analysis of the biofilm exopolymeric matrix carbohydrate composition of corrosion-aggressive bacteria].

    PubMed

    Purish, L M; Asaulenko, L G; Abdulina, D R; Voĭtchuk, S I; Iutinskaia, G A

    2013-01-01

    The carbohydrate components of biofilms of corrosion-aggressive bacteria were studied by transmisstion electron microscopy using lectins labeled with colloidal gold. N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and neutral carbohydrates D-glucose and D-mannose were found within the exopolymeric matrix. Lectins with equal carbohydrate specificity demonstrated different degrees of interaction with the carbohydrate components of bacterial biofilms. To identify N-acetyl-D-galactosamine in biofilms of Desulfovibrio sp. 10 and Bacillus subtilis 36, the LBA lectin appeared to be most specific; in the case of N-acetyl-D-glucosamine in biofilms of B. subtilis 36 and Pseudomonas aeruginosa 27, the WGA lectin. During visualization of neutral carbohydrates in the studied cultures, the PSA lectin was most specific. We have shown that lectins labeled with colloidal gold could be used as an express method for the identification and localization of carbohydrates in glycopolymers of the biofilm exopolymeric matrix.

  16. Lectin Activation in Giardia lamblia by Host Protease: A Novel Host-Parasite Interaction

    NASA Astrophysics Data System (ADS)

    Lev, Boaz; Ward, Honorine; Keusch, Gerald T.; Pereira, Miercio E. A.

    1986-04-01

    A lectin in Giardia lamblia was activated by secretions from the human duodenum, the environment where the parasite lives. Incubation of the secretions with trypsin inhibitors prevented the appearance of lectin activity, implicating proteases as the activating agent. Accordingly, lectin activation was also produced by crystalline trypsin and Pronase; other proteases tested were ineffective. When activated, the lectin agglutinated intestinal cells to which the parasite adheres in vivo. The lectin was most specific to mannose-6-phosphate and apparently was bound to the plasma membrane. Activation of a parasite lectin by a host protease represents a novel mechanism of hostparasite interaction and may contribute to the affinity of Giardia lamblia to the infection site.

  17. Could plant lectins become promising anti-tumour drugs for causing autophagic cell death?

    PubMed

    Liu, Z; Luo, Y; Zhou, T-T; Zhang, W-Z

    2013-10-01

    Plant lectins, a group of highly diverse carbohydrate-binding proteins of non-immune origin, are ubiquitously distributed through a variety of plant species, and have recently drawn rising attention due to their remarkable ability to kill tumour cells using mechanisms implicated in autophagy. In this review, we provide a brief outline of structures of some representative plant lectins such as concanavalin A, Polygonatum cyrtonema lectin and mistletoe lectins. These can target autophagy by modulating BNIP-3, ROS-p38-p53, Ras-Raf and PI3KCI-Akt pathways, as well as Beclin-1, in many types of cancer cells. In addition, we further discuss how plant lectins are able to kill cancer cells by modulating autophagic death, for therapeutic purposes. Together, these findings provide a comprehensive perspective concerning plant lectins as promising new anti-tumour drugs, with respect to autophagic cell death in future cancer therapeutics.

  18. Leguminous lectins as tools for studying the role of sugar residues in leukocyte recruitment.

    PubMed Central

    Alencar, N M; Teixeira, E H; Assreuy, A M; Cavada, B S; Flores, C A; Ribeiro, R A

    1999-01-01

    The natural physiological ligands for selectins are oligosaccharides found in glycoprotein or glycolipid molecules in cell membranes. In order to study the role of sugar residues in the in vivo lectin anti-inflammatory effect, we tested three leguminous lectins with different carbohydrate binding affinities in the peritonitis and paw oedema models induced by carrageenin in rats. L. sericeus lectin was more anti-inflammatory than D. virgata lectin, the effects being reversed by their specific binding sugars (N-acetylglucosamine and alpha-methylmannoside, respectively). However, V. macrocarpa, a galactose-specific lectin, was not anti-inflammatory. The proposed anti-inflammatory activity of lectins could be due to a blockage of neutrophil-selectin carbohydrate ligands. Thus, according to the present data, we suggest an important role for N-acetylglucosamine residue as the major ligand for selectins on rat neutrophil membranes. PMID:10704148

  19. Activation of phenoloxidase activity by humoral lectin in hemocytes of freshwater crab Paratelphusa jacquemontii.

    PubMed

    Denis, Maghil; Ramasamy, Sivakumar Mullaivanam; Kamalanathan, Tamilarasan; Thayappan, Karthigayani; Mannarreddy, Prabu; Doss, Bhuvaneswari Selvaputhiran; Munusamy, Arumugam

    2017-04-01

    The lectin, Pjlec isolated from the hemolymph of the freshwater crab Paratelphusa jacquemontii hemagglutinated (HA) with mice, rabbit and rat erythrocytes. However, the lectin failed to agglutinate neraminidase treated asialylated erythrocytes showing its sialic acid specificity. The poyacyrlamide gel electrophoresis of lectin yielded 310kDa proteins, on sodium sulphate dodecyl (SDS) gel appeared as a tetramer with subunits of 76kDa. The observation of in vitro phagocytosis in granular hemocytes of lectin opsonized rabbit erythrocyte by Transmission electron microscopy (TEM) showed the release of lytic vesicles by exocytosis prior to engulfment. The Pjlec lectin also showed an ability to oxidize L-3, 4 dihydroxyphenylalanine (L-DOPA) and in hemocyte lysate preparation (HLS) was enhanced on reduction with SDS and on proteolytic cleavage with trypsin. The lectin appeared to have a regulatory role in activation of enzyme activity associated with phagocytosis and melanin formation. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Development and Applications of Lectins as Biological Tools in Biomedical Research.

    PubMed

    Dan, Xiuli; Liu, Wenlong; Ng, Tzi Bun

    2016-03-01

    As a new and burgeoning area following genomics and proteomics, glycomics has become a hot issue due to its pivotal roles in many physiological and pathological processes. Glycans are much more complicated than genes or proteins since glycans are highly branched and dynamic. Antibodies and lectins are the two major molecular tools applied for glycan profiling. Though the study of antibodies and lectins started at almost the same time in 1880s, lectins gained much less attention than the antibodies until recent decades when the importance and difficulties of glycomics were realized. The present review summarizes the discovery history of lectins and their biological functions with a special emphasis on their various applications as biological tools. Both older techniques that had been developed in the last century and new technologies developed in recent years, especially lectin microarrays and lectin-based biosensors, are included in this account. © 2015 Wiley Periodicals, Inc.

  1. Isolation of the galactose-binding lectin that mediates the in vitro adherence of Entamoeba histolytica.

    PubMed Central

    Petri, W A; Smith, R D; Schlesinger, P H; Murphy, C F; Ravdin, J I

    1987-01-01

    Entamoeba histolytica adheres to human colonic mucus, colonic epithelial cells, and other target cells via a galactose (Gal) or N-acetyl-D-galactosamine (GalNAc) inhibitable surface lectin. Blockade of this adherence lectin with Gal or GalNAc in vitro prevents amebic killing of target cells. We have identified and purified the adherence lectin by two methods: affinity columns derivatized with galactose monomers or galactose terminal glycoproteins, and affinity columns and immunoblots prepared with monoclonal antibodies that inhibit amebic adherence. By both methods the adherence lectin was identified as a 170-kD secreted and membrane-bound amebic protein. The surface location of the lectin was confirmed by indirect immunofluorescence. Purified lectin competitively inhibited amebic adherence to target cells by binding to receptors on the target Chinese hamster ovary cells in a Gal-inhibitable manner. Images PMID:2890654

  2. A galactose-specific lectin from the hemolymph of the pearl oyster, Pinctada fucata martensii.

    PubMed

    Suzuki, T; Mori, K

    1989-01-01

    1. A lectin in the serum of Pinctada fucata martensii was purified by a combination of affinity chromatography on Sepharose 4B coupled with bovine submaxillary gland mucine, anion exchange chromatography on Mono Q and gel filtration on Superose 6. 2. The purified lectin was indicated to be homogeneous by polyacrylamide electrophoresis and rechromatography on Mono Q. 3. The purified lectin was approximately 440,000 in molecular weight and was composed of identical subunits with a molecular weight of approximately 20,000. 4. D-galactose and N-acetylgalactosamine gave a 50% inhibition of agglutination of horse erythrocytes by the lectin at 0.3 and 1.2 mM, respectively. 5. The antibody obtained from rabbit immunized with the purified lectin was monospecific to the lectin judged from the hemagglutination blocking test, immunoelectrophoresis and immunoblotting.

  3. Pharmacological inhibition of mannose-binding lectin ameliorates neurobehavioral dysfunction following experimental traumatic brain injury.

    PubMed

    De Blasio, Daiana; Fumagalli, Stefano; Longhi, Luca; Orsini, Franca; Palmioli, Alessandro; Stravalaci, Matteo; Vegliante, Gloria; Zanier, Elisa R; Bernardi, Anna; Gobbi, Marco; De Simoni, Maria-Grazia

    2017-03-01

    Mannose-binding lectin is present in the contusion area of traumatic brain-injured patients and in that of traumatic brain-injured mice, where mannose-binding lectin-C exceeds mannose-binding lectin-A. The reduced susceptibility to traumatic brain injury of mannose-binding lectin double knock-out mice (mannose-binding lectin(-/-)) when compared to wild type mice suggests that mannose-binding lectin may be a therapeutic target following traumatic brain injury. Here, we evaluated the effects of a multivalent glycomimetic mannose-binding lectin ligand, Polyman9, following traumatic brain injury in mice. In vitro surface plasmon resonance assay indicated that Polyman9 dose-dependently inhibits the binding to immobilized mannose residues of plasma mannose-binding lectin-C selectively over that of mannose-binding lectin-A. Male C57Bl/6 mice underwent sham/controlled cortical impact traumatic brain injury and intravenous treatment with Polyman9/saline. Ex-vivo surface plasmon resonance studies confirmed that Polyman9 effectively reduces the binding of plasma mannose-binding lectin-C to immobilized mannose residues. In vivo studies up to four weeks post injury, showed that Polyman9 induces significant improvement in sensorimotor deficits (by neuroscore and beam walk), promotes neurogenesis (73% increase in doublecortin immunoreactivity), and astrogliosis (28% increase in glial fibrillary acid protein). Polyman9 administration in brain-injured mannose-binding lectin(-/-) mice had no effect on post-traumatic brain-injured functional deficits, suggestive of the specificity of its neuroprotective effects. The neurobehavioral efficacy of Polyman9 implicates mannose-binding lectin-C as a novel therapeutic target for traumatic brain injury.

  4. Unique posttranslational modifications of chitin-binding lectins of Entamoeba invadens cyst walls.

    PubMed

    Van Dellen, Katrina L; Chatterjee, Anirban; Ratner, Daniel M; Magnelli, Paula E; Cipollo, John F; Steffen, Martin; Robbins, Phillips W; Samuelson, John

    2006-05-01

    Entamoeba histolytica, which causes amebic dysentery and liver abscesses, is spread via chitin-walled cysts. The most abundant protein in the cyst wall of Entamoeba invadens, a model for amebic encystation, is a lectin called EiJacob1. EiJacob1 has five tandemly arrayed, six-Cys chitin-binding domains separated by low-complexity Ser- and Thr-rich spacers. E. histolytica also has numerous predicted Jessie lectins and chitinases, which contain a single, N-terminal eight-Cys chitin-binding domain. We hypothesized that E. invadens cyst walls are composed entirely of proteins with six-Cys or eight-Cys chitin-binding domains and that some of these proteins contain sugars. E. invadens genomic sequences predicted seven Jacob lectins, five Jessie lectins, and three chitinases. Reverse transcription-PCR analysis showed that mRNAs encoding Jacobs, Jessies, and chitinases are increased during E. invadens encystation, while mass spectrometry showed that the cyst wall is composed of an approximately 30:70 mix of Jacob lectins (cross-linking proteins) and Jessie and chitinase lectins (possible enzymes). Three Jacob lectins were cleaved prior to Lys at conserved sites (e.g., TPSVDK) in the Ser- and Thr-rich spacers between chitin-binding domains. A model peptide was cleaved at the same site by papain and E. invadens Cys proteases, suggesting that the latter cleave Jacob lectins in vivo. Some Jacob lectins had O-phosphodiester-linked carbohydrates, which were one to seven hexoses long and had deoxysugars at reducing ends. We concluded that the major protein components of the E. invadens cyst wall all contain chitin-binding domains (chitinases, Jessie lectins, and Jacob lectins) and that the Jacob lectins are differentially modified by site-specific Cys proteases and O-phosphodiester-linked glycans.

  5. Unique Posttranslational Modifications of Chitin-Binding Lectins of Entamoeba invadens Cyst Walls

    PubMed Central

    Van Dellen, Katrina L.; Chatterjee, Anirban; Ratner, Daniel M.; Magnelli, Paula E.; Cipollo, John F.; Steffen, Martin; Robbins, Phillips W.; Samuelson, John

    2006-01-01

    Entamoeba histolytica, which causes amebic dysentery and liver abscesses, is spread via chitin-walled cysts. The most abundant protein in the cyst wall of Entamoeba invadens, a model for amebic encystation, is a lectin called EiJacob1. EiJacob1 has five tandemly arrayed, six-Cys chitin-binding domains separated by low-complexity Ser- and Thr-rich spacers. E. histolytica also has numerous predicted Jessie lectins and chitinases, which contain a single, N-terminal eight-Cys chitin-binding domain. We hypothesized that E. invadens cyst walls are composed entirely of proteins with six-Cys or eight-Cys chitin-binding domains and that some of these proteins contain sugars. E. invadens genomic sequences predicted seven Jacob lectins, five Jessie lectins, and three chitinases. Reverse transcription-PCR analysis showed that mRNAs encoding Jacobs, Jessies, and chitinases are increased during E. invadens encystation, while mass spectrometry showed that the cyst wall is composed of an ∼30:70 mix of Jacob lectins (cross-linking proteins) and Jessie and chitinase lectins (possible enzymes). Three Jacob lectins were cleaved prior to Lys at conserved sites (e.g., TPSVDK) in the Ser- and Thr-rich spacers between chitin-binding domains. A model peptide was cleaved at the same site by papain and E. invadens Cys proteases, suggesting that the latter cleave Jacob lectins in vivo. Some Jacob lectins had O-phosphodiester-linked carbohydrates, which were one to seven hexoses long and had deoxysugars at reducing ends. We concluded that the major protein components of the E. invadens cyst wall all contain chitin-binding domains (chitinases, Jessie lectins, and Jacob lectins) and that the Jacob lectins are differentially modified by site-specific Cys proteases and O-phosphodiester-linked glycans. PMID:16682461

  6. Isolation and partial characterization of a lectin from a false brome grass (Brachypodium sylvaticum).

    PubMed Central

    Peumans, W J; Spaepen, C; Stinissen, H M; Carlier, A R

    1982-01-01

    A lectin has been isolated from embryos of a false brome grass species (Brachypodium sylvaticum) by affinity chromatography on immobilized N-acetylglucosamine. It is a dimeric protein of two identical subunits of mol.wt. 18 000. Although it resembles cereal lectins with respect to its biochemical and physicochemical properties, it differs structurally in several aspects from wheat-germ-agglutinin-like lectins. Images Fig. 1. Fig. 3. Fig. 4. PMID:6816219

  7. [Lectins and the problem of phitopatogene recognition by a host plant].

    PubMed

    Babosha, A V

    2008-01-01

    Under consideration are some questions concerning participation of lectins in the plant pathogenesis, including their role in the recognition of microbes and elicitors, and as a protective agent limiting pathogenic growth and displacements. "Classical" lectins also probably play an important role in these processes along with lectin-like receptor kinases. The principal features of those "classical" lectins are their relativly high concentration in the plant tissues, monosaccharide specificity, and limited number of the isolecin forms. Therefore, in supposing their participation in the biological recognition, it is needed to clarify how does a limited number of lectins with a limited number of carbohydrate groups can provide recognition of a potentially huge number of pathogens. This task can be fulfilled by recognition of carbohydrate residues peculiar to a particular microbe group by the "classical" lectins. These recognition processes are similar to acivity of the animal inherited immune system responsible for a rapid primary protection even in animals with well developed antibody system. A mechanism widening the carbohydrate specificity of the carbohydrate-binding center includes interaction with hydrophobic substituents in a carbohydrate residue, as well as lectin modular organization allowing for regulation of lectin binding with oligo- and polysaccharides. The free lectins effect on the microbe growth in both plants and animals. Such an action may be inhibiting in pathogenesis, while in the case of symbiotic relations, the lectin can bear signal that readdresses metabolism of a future symbiont. So, lectins seem to serve as natural deciphering device for information contained in the carbohydrate polymers, and reading of this information is the main lectin function in the cell.

  8. Structural and functional overview of the lectin complement pathway: its molecular basis and physiological implication.

    PubMed

    Matsushita, Misao; Endo, Yuichi; Fujita, Teizo

    2013-08-01

    The complement system is an effector mechanism in immunity. It is activated in three ways, the classical, alternative and lectin pathways. The lectin pathway is initiated by the binding of mannose-binding lectin (MBL) or ficolins to carbohydrates on the surfaces of pathogens. In humans, MBL and three types of ficolins (L-ficolin, H-ficolin, and M-ficolin) are present in plasma. Of these lectins, at least, MBL, L-ficolin, and H-ficolin are complexed with three types of MBL-associated serine proteases (MASPs), MASP-1, MASP-2, and MASP-3 and their truncated proteins (MAp44 and sMAP). In the lectin pathway, the lectin-MASP complex (i.e., a complex of lectin, MASPs and their truncated proteins) binds to pathogens, resulting in the activation of C4 and C2 to generate a C3 convertase capable of activating C3. MASP-2 is involved in the activation of C4 and C2. MASP-1 activates C2 and MASP-2. The functions of MASP-3, sMAP, and MAp44 in the lectin pathway remain unknown. MASP-1 and MASP-3 also have a role in the alternative pathway. MBL and ficolins are able to bind to a variety of pathogens depending on their carbohydrate binding specificity, resulting in the activation of the lectin pathway. Deficiencies of the components of the lectin pathway are associated to susceptibility to infection, indicating an important role of the lectin pathway in innate immunity. The lectin-MASP complex is also involved in innate immunity by activating the coagulation system. Recent findings suggest a crucial role of MASP-3 in development.

  9. Large Scale Magnetic Separation of Solanum tuberosum Tuber Lectin from Potato Starch Waste Water

    NASA Astrophysics Data System (ADS)

    Safarik, Ivo; Horska, Katerina; Martinez, Lluis M.; Safarikova, Mirka

    2010-12-01

    A simple procedure for large scale isolation of Solanum tuberosum tuber lectin from potato starch industry waste water has been developed. The procedure employed magnetic chitosan microparticles as an affinity adsorbent. Magnetic separation was performed in a flow-through magnetic separation system. The adsorbed lectin was eluted with glycine/HCl buffer, pH 2.2. The specific activity of separated lectin increased approximately 27 times during the isolation process.

  10. Stability, subunit interactions and carbohydrate-binding of the seed lectin from Pterocarpus angolensis.

    PubMed

    Echemendia-Blanco, Dannele; Van Driessche, Edilbert; Ncube, Ignatious; Read, John S; Beeckmans, Sonia

    2009-01-01

    From 1 kg of defatted Pterocarpus angolensis (mukwa tree) seed meal, 21.6 grams of an alpha,D-mannose/glucose-specific lectin can be purified on mannose-Sepharose. Relative affinities for several (oligo)saccharides and glycoproteins were studied by haemagglutination-inhibition. Gel filtration shows that the lectin exists as a dimer above pH 5 and as a monomer below pH 3.5. This is confirmed by studies on the release of lectin subunits that were adsorbed from solution to lectin monomers immobilized onto Eupergit-c. From the gel filtration patterns it is calculated that a residue with pK(a) of about 4.4 is involved in dimer dissociation. Titration of glutamic acids (E60, E209) is postulated to be involved. CD spectroscopy shows that the secondary structure of the lectin is unchanged between pH 1 and 12.5, and that the tertiary structure remains unchanged between pH 5 and 12. In the acid pH region, reversible spectral changes occur that may be due to the titration of one or more amino acids with a pK(a) value of 3.9-4.2, probably aspartic acid. These residues are implicated in sugar-binding but not in dimerization of the lectin. Only at pH 12.5, irreversible denaturation occurs. Mukwa lectin displays full carbohydrate-binding capacity between pH 4 and 12, as is concluded from ELLA (Enzyme Linked Lectin Assay) using ovalbumin and fetuin, and from binding of the same glycoproteins to immobilized lectin monomers. The lectin is rapidly and fully reversibly demetallized at pH 2.5 with 5 mM EDTA. The demetallized lectin is completely devoid of sugar-binding activity. Mukwa lectin is a very thermostable molecule (at least till 85 degrees C). However, addition of non-ionic detergents substantially lowers its thermostability.

  11. Intracellular lectins are involved in quality control of glycoproteins

    PubMed Central

    YAMAMOTO, Kazuo

    2014-01-01

    Glycoprotein quality control is categorized into three kinds of reactions; the folding of nascent glycoproteins, ER-associated degradation of misfolded or unassembled glycoproteins, and transport and sorting of correctly folded glycoproteins. In all three processes, N-glycans on the glycoproteins are used as tags that are recognized by intracellular lectins. We analyzed the functions of these intracellular lectins and their sugar-binding specificities. The results clearly showed that the A, B, and C-arms of high mannose-type glycans participate in the folding, transport and sorting, and degradation, respectively, of newly synthesized peptides. After correctly folded glycoproteins are transported to the Golgi apparatus, N-glycans are trimmed into Man3GlcNAc2 and then rebuilt into various complex-type glycans in the Golgi, resulting in the addition of diverse sugar structures that allow glycoproteins to play various roles outside of the cells. PMID:24522156

  12. The role of lectins in allergic sensitization and allergic disease.

    PubMed

    Salazar, Fabián; Sewell, Herb F; Shakib, Farouk; Ghaemmaghami, Amir M

    2013-07-01

    Allergic diseases are a global public health issue affecting millions of persons around the world. However, full understanding of the molecular basis of this group of chronic inflammatory disorders remains rather elusive. Recently, the role of carbohydrates on allergens and their counterstructures on antigen-presenting cells (lectins) have been highlighted as crucial factors in allergen sensitization, which culminates in TH2 cell differentiation and the production of deleterious specific IgE antibodies. Here we review recent progress on the role of different lectins in patients with type I hypersensitivity or allergy, their interplay with other determinants of allergenicity, and ways of developing therapeutic modalities against newly identified targets. Copyright © 2013 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  13. Plant Lectins as Medical Tools against Digestive System Cancers.

    PubMed

    Estrada-Martínez, Laura Elena; Moreno-Celis, Ulisses; Cervantes-Jiménez, Ricardo; Ferriz-Martínez, Roberto Augusto; Blanco-Labra, Alejandro; García-Gasca, Teresa

    2017-07-03

    Digestive system cancers-those of the esophagus, stomach, small intestine, colon-rectum, liver, and pancreas-are highly related to genetics and lifestyle. Most are considered highly mortal due to the frequency of late diagnosis, usually in advanced stages, caused by the absence of symptoms or masked by other pathologies. Different tools are being investigated in the search of a more precise diagnosis and treatment. Plant lectins have been studied because of their ability to recognize and bind to carbohydrates, exerting a variety of biological activities on animal cells, including anticancer activities. The present report integrates existing information on the activity of plant lectins on various types of digestive system cancers, and surveys the current state of research into their properties for diagnosis and selective treatment.

  14. Purification and biological effects of Araucaria angustifolia (Araucariaceae) seed lectin

    SciTech Connect

    Santi-Gadelha, Tatiane; Almeida Gadelha, Carlos Alberto de; Aragao, Karoline Saboia; Gomes, Raphaela Cardoso; Freitas Pires, Alana de; Toyama, Marcos Hikari; Oliveira Toyama, Daniela de; Nunes de Alencar, Nylane Maria; Criddle, David Neil; Assreuy, Ana Maria Sampaio . E-mail: assreuy@uece.br; Cavada, Benildo Sousa . E-mail: bscavada@ufc.br

    2006-12-01

    This paper describes the purification and characterization of a new N-acetyl-D-glucosamine-specific lectin from Araucaria angustifolia (AaL) seeds (Araucariaceae) and its anti-inflammatory and antibacterial activities. AaL was purified using a combination of affinity chromatography on a chitin column and ion exchange chromatography on Sephacel-DEAE. The pure protein has 8.0 kDa (SDS-PAGE) and specifically agglutinates rabbit erythrocytes, effect that was independent of the presence of divalent cations and was inhibited after incubation with glucose and N-acetyl-D-glucosamine. AaL showed antibacterial activity against Gram-negative and Gram-positive strains, shown by scanning electron microscopy. AaL, intravenously injected into rats, showed anti-inflammatory effect, via carbohydrate site interaction, in the models of paw edema and peritonitis. This lectin can be used as a tool for studying bacterial infections and inflammatory processes.

  15. Griffithsin: An Antiviral Lectin with Outstanding Therapeutic Potential

    PubMed Central

    Lusvarghi, Sabrina; Bewley, Carole A.

    2016-01-01

    Griffithsin (GRFT), an algae-derived lectin, is one of the most potent viral entry inhibitors discovered to date. It is currently being developed as a microbicide with broad-spectrum activity against several enveloped viruses. GRFT can inhibit human immunodeficiency virus (HIV) infection at picomolar concentrations, surpassing the ability of most anti-HIV agents. The potential to inhibit other viruses as well as parasites has also been demonstrated. Griffithsin’s antiviral activity stems from its ability to bind terminal mannoses present in high-mannose oligosaccharides and crosslink these glycans on the surface of the viral envelope glycoproteins. Here, we review structural and biochemical studies that established mode of action and facilitated construction of GRFT analogs, mechanisms that may lead to resistance, and in vitro and pre-clinical results that support the therapeutic potential of this lectin. PMID:27783038

  16. Gliadins bind to reticulin in a lectin-like manner.

    PubMed

    Unsworth, D J; Leonard, J N; Hobday, C M; Griffiths, C E; Powles, A V; Haffenden, G P; Fry, L

    1987-01-01

    It has previously been reported that gliadins bind to reticulin in tissue sections. Three lines of evidence are reported in this study which indicate that the gliadins bind to reticulins because they are lectins which bind to sugars expressed on glycoproteins in reticulin and other sites. First, immunofluorescence studies on tissue sections showed that although gliadin binding is largely confined to areas rich in reticulin, it is, nonetheless, also seen in one or two other sites devoid of reticulin. Second, by using fluorescein-labelled lectins of known specificity, it has been shown that the areas to which gliadins bind in tissue sections (including those sites devoid of reticulin) are rich in particular sugars. Third, it has been shown that one of these sugars, alpha-D-mannose, partially inhibited gliadin binding to tissue sections.

  17. C-type lectins in immunity: recent developments

    PubMed Central

    Dambuza, Ivy M; Brown, Gordon D

    2015-01-01

    C-type lectin receptors (CLRs) comprise a large superfamily of proteins, which recognise a diverse range of ligands, and are defined by the presence of at least one C-type lectin-like domain (CTLD). Of particular interest are the single extracellular CTLD-containing receptors of the ‘Dectin-1’ and ‘Dectin-2’ clusters, which associate with signalling adaptors or possess integral intracellular signalling domains. These CLRs have traditionally been associated with the recognition of fungi, but recent discoveries have revealed diverse and unexpected functions. In this review, we describe their newly identified roles in anti-microbial host defence, homeostasis, autoimmunity, allergy and their functions in the recognition and response to dead and cancerous cells. PMID:25553393

  18. Parallel quantification of lectin-glycan interaction using ultrafiltration.

    PubMed

    Takeda, Yoichi; Seko, Akira; Sakono, Masafumi; Hachisu, Masakazu; Koizumi, Akihiko; Fujikawa, Kohki; Ito, Yukishige

    2013-06-28

    Using ultrafiltration membrane, a simple method for screening protein-ligand interaction was developed. The procedure comprises three steps: mixing ligand with protein, ultrafiltration of the solution, and quantification of unbound ligands by HPLC. By conducting analysis with variable protein concentrations, affinity constants were easily obtained. Multiple ligands can be analyzed simultaneously as a mixture, when concentration of ligands was controlled. Feasibility of this method for lectin-glycan interaction analysis was examined using fluorescently labeled high-mannose-type glycans and recombinant intracellular lectins or endo-α-mannosidase mutants. Estimated Ka values of malectin and VIP36 were in good agreement indeed with those evaluated by conventional methods such as isothermal titration calorimetry (ITC) or frontal affinity chromatography (FAC). Finally, several mutants of endo-α-mannosidase were produced and their affinities to monoglucosylated glycans were evaluated.

  19. Recombinant chimeric lectins consisting of mannose-binding lectin and L-ficolin are potent inhibitors of influenza A virus compared with mannose-binding lectin

    PubMed Central

    Chang, Wei-Chuan; Hartshorn, Kevan L.; White, Mitchell R.; Moyoa, Patience; Michelow, Ian C.; Koziel, Henry; Kinane, Bernard T.; Schmidt, Emmett V.; Fujita, Teizo; Takahashi, Kazue

    2010-01-01

    MBL structurally contains a type II-like collagenous domain and a carbohydrate recognition domain (CRD). We have recently generated three novel recombinant chimeric lectins (RCL), in which varying length of collagenous domain of mannose-binding lectin (MBL) is replaced with that of L-ficolin (L-FCN). CRD of MBL is used for target recognition because it has a broad spectrum in pathogen recognition compared with L-FCN. Results of our study demonstrate that these RCLs are potent inhibitors of influenza A virus (IAV). RCLs, against IAV, show dose-dependent activation of the lectin complement pathway, which is significantly higher than that of recombinant human MBL (rMBL). This activity is observed even without MBL-associated serine proteases (MASPs, provided by MBL deficient mouse sera), which have been thought to mediate complement activation. These observations suggest that RCLs are more efficient in associating with MASP-2, which predominantly mediates the activity. Yet, additional serum further increases the activity while RCL-mediated coagulation-like enzyme activities are diminished compared with rMBL, suggesting reduced association with MASP-1, which has been shown to mediate coagulation-like activity. These data suggest that RCLs may interfere less with host coagulation, which is advantageous to be a therapeutic drug. Importantly, these RCLs have surpassed rMBL for anti-viral activities, such as viral aggregation, reduction of viral hemagglutination (HA) and inhibition of virus-mediated HA and neuraminidase (NA) activities. These results are encouraging that novel RCLs could be used as anti-IAV agents with less side effect and that RCLs would be suitable candidates in developing a new anti-IAV therapy. PMID:21035429

  20. Recombinant chimeric lectins consisting of mannose-binding lectin and L-ficolin are potent inhibitors of influenza A virus compared with mannose-binding lectin.

    PubMed

    Chang, Wei-Chuan; Hartshorn, Kevan L; White, Mitchell R; Moyo, Patience; Michelow, Ian C; Koziel, Henry; Kinane, Bernard T; Schmidt, Emmett V; Fujita, Teizo; Takahashi, Kazue

    2011-02-01

    MBL structurally contains a type II-like collagenous domain and a carbohydrate recognition domain (CRD). We have recently generated three novel recombinant chimeric lectins (RCL), in which varying length of collagenous domain of mannose-binding lectin (MBL) is replaced with that of L-ficolin (L-FCN). CRD of MBL is used for target recognition because it has a broad spectrum in pathogen recognition compared with L-FCN. Results of our study demonstrate that these RCLs are potent inhibitors of influenza A virus (IAV). RCLs, against IAV, show dose-dependent activation of the lectin complement pathway, which is significantly higher than that of recombinant human MBL (rMBL). This activity is observed even without MBL-associated serine proteases (MASPs, provided by MBL deficient mouse sera), which have been thought to mediate complement activation. These observations suggest that RCLs are more efficient in associating with MASP-2, which predominantly mediates the activity. Yet, additional serum further increases the activity while RCL-mediated coagulation-like enzyme activities are diminished compared with rMBL, suggesting reduced association with MASP-1, which has been shown to mediate coagulation-like activity. These data suggest that RCLs may interfere less with host coagulation, which is advantageous to be a therapeutic drug. Importantly, these RCLs have surpassed rMBL for anti-viral activities, such as viral aggregation, reduction of viral hemagglutination (HA) and inhibition of virus-mediated HA and neuraminidase (NA) activities. These results are encouraging that novel RCLs could be used as anti-IAV agents with less side effect and that RCLs would be suitable candidates in developing a new anti-IAV therapy. Copyright © 2010 Elsevier Inc. All rights reserved.

  1. Mannan-Binding Lectin Enhances Susceptibility to Visceral Leishmaniasis

    PubMed Central

    de Miranda Santos, Isabel K. F.; Costa, Carlos H. N.; Krieger, Henrique; Feitosa, Mary F.; Zurakowski, David; Fardin, Babak; Gomes, Regis B. B.; Weiner, Debra L.; Harn, Donald A.; Ezekowitz, R. Alan B.; Epstein, Judith E.

    2001-01-01

    Levels of the serum opsonin mannan-binding lectin (MBL) were directly correlated with the probability of developing visceral leishmaniasis. Monocytes infected with MBL-opsonized Leishmania chagasi promastigotes secreted higher levels of tumor necrosis factor alpha and interleukin-6 than cells infected with nonopsonized parasites. Our findings indicate that MBL can modulate the clinical outcome of infection with L. chagasi and the function of infected macrophages. PMID:11447210

  2. The Lectin Pathway of Complement and Rheumatic Heart Disease

    PubMed Central

    Beltrame, Marcia Holsbach; Catarino, Sandra Jeremias; Goeldner, Isabela; Boldt, Angelica Beate Winter; de Messias-Reason, Iara José

    2014-01-01

    The innate immune system is the first line of host defense against infection and is comprised of humoral and cellular mechanisms that recognize potential pathogens within minutes or hours of entry. The effector components of innate immunity include epithelial barriers, phagocytes, and natural killer cells, as well as cytokines and the complement system. Complement plays an important role in the immediate response against microorganisms, including Streptococcus sp. The lectin pathway is one of three pathways by which the complement system can be activated. This pathway is initiated by the binding of mannose-binding lectin (MBL), collectin 11 (CL-K1), and ficolins (Ficolin-1, Ficolin-2, and Ficolin-3) to microbial surface oligosaccharides and acetylated residues, respectively. Upon binding to target molecules, MBL, CL-K1, and ficolins form complexes with MBL-associated serine proteases 1 and 2 (MASP-1 and MASP-2), which cleave C4 and C2 forming the C3 convertase (C4b2a). Subsequent activation of complement cascade leads to opsonization, phagocytosis, and lysis of target microorganisms through the formation of the membrane-attack complex. In addition, activation of complement may induce several inflammatory effects, such as expression of adhesion molecules, chemotaxis and activation of leukocytes, release of reactive oxygen species, and secretion of cytokines and chemokines. In this chapter, we review the general aspects of the structure, function, and genetic polymorphism of lectin-pathway components and discuss most recent understanding on the role of the lectin pathway in the predisposition and clinical progression of Rheumatic Fever. PMID:25654073

  3. Genotoxicity evaluation of Moringa oleifera seed extract and lectin.

    PubMed

    Rolim, Lucíola A D M M; Macêdo, Márcia F S; Sisenando, Herbert Ary; Napoleão, Thiago H; Felzenszwalb, Israel; Aiub, Claudia A F; Coelho, Luana C B B; Medeiros, Silvia R B; Paiva, Patrícia M G

    2011-03-01

    This article reports the genotoxicity assessment of an extract of M. oleifera seed powder and the water-soluble Moringa oleifera lectin (WSMoL) isolated from seeds. The lectin isolated by chitin chromatography showed hemagglutinating activity with different erythrocytes, activity in a broad pH range (4.5 to 9.5), and retention of hemagglutinating activity after being heated to 100 °C. Genotoxicity of the seed extract and WSMoL were assessed using the cell-free plasmid DNA as well as the Salmonella typhimurium (Ames and Kado) assays with TA97, TA98, TA100, and TA102 in the presence or absence of hepatic metabolization. Seed extract at concentration (0.2 μg/μL) recommended to treat water was not genotoxic by Ames, Kado, and cell-free plasmid DNA assays. S. typhimurium strains showed to be sensitive to M. oleifera extract revealing a mutagenic effect at doses higher than 0.6 μg/μL with hepatic metabolization. The extract at doses higher than 0.4 μg/μL, without hepatic metabolization, was mutagenic for TA100 and TA102. WSMoL was nonmutagenic by used assays. The use of high concentrations of the extract may pose a risk to human health and the safe use of M. oleifera seed powder to treat water for human consumption requires more study; however, the purified lectin could be an alternative for water treatment. The concentration 0.2 μg/μL of M. oleifera seed extract recommended to treat water for humans did not pose a risk to human health. The mutagenicity detected at concentrations higher than 0.4 μg/μL was not due to WSMoL, lectin isolated from extract.

  4. Factors affecting binding of galacto ligands to Actinomyces viscosus lectin.

    PubMed Central

    Heeb, M J; Marini, A M; Gabriel, O

    1985-01-01

    The specificity requirements for the binding of Actinomyces viscosus T14V were examined by testing simple sugars, oligopeptides, and glycoproteins as inhibitors of the aggregation of glycoprotein-coated latex beads and washed A. viscosus cells. Lactose was the most inhibitory simple sugar; D-fucose and D-galactose were equally inhibitory, methyl-alpha-D-fucoside was slightly less inhibitory, and L-fucose and raffinose were not inhibitory. The concentration of galactose residues required for 50% inhibition of aggregation was 15 times higher in the form of lactose than in the form of asialoglycoprotein, suggesting an enhancement of lectin binding when galactose residues are clustered. However, when the inhibitory power of bi-, tri-, and tetraantennary asialooligopeptides of alpha 1-acid glycoprotein was compared with that of equivalent concentrations of galactose in the form of lactose, the biantennary form was slightly less effective than lactose, the triantennary form was approximately as effective as lactose, and the tetraantennary form was slightly more effective than lactose. Steric interference may prevent this type of clustering from enhancing lectin binding. The O-linked asialooligopeptides of asialofetuin were 10 times more inhibitory than an equivalent concentration of galactose in the form of N-linked asialooligopeptides. Thus, galactose beta-1----3 linked to N-acetylgalactosamine exhibits greater specificity for the A. viscosus lectin than does galactose beta-1----4 linked to N-acetylglucosamine. These results, taken together with previously reported data, are consistent with a lectin of low affinity, binding enhanced by multivalency, and specificity for beta-linked galactose. PMID:2578122

  5. A glycobiology review: carbohydrates, lectins, and implications in cancer therapeutics

    PubMed Central

    Ghazarian, Haike; Idoni, Brian; Oppenheimer, Steven B.

    2010-01-01

    This review is intended for general readers who would like a basic foundation in carbohydrate structure and function, lectin biology and the implications of glycobiology in human health and disease, particularly in cancer therapeutics. These topics are among the hundreds included in the field of glycobiology and are treated here because they form the cornerstone of glycobiology or the focus of many advances in this rapidly expanding field. PMID:20199800

  6. Lectins as membrane components of mitochondria from Ricinus communis.

    PubMed Central

    Bowles, D J; Schnarrenberger, C; Kauss, H

    1976-01-01

    1. Mitochondria were isolated from developing endosperm of Ricinus communis and were fractionated into outer membrane and inner membrane. The relative purity of the two membrane fractions was determined by marker enzymes. The fractions were also examined by negative-stain electron microscopy. 2. Membrane fractions were sequentially extracted in the following way. (a) Suspension in 0.5M-potassium phosphate, pH7.1; (b)suspension in 0.1M-EDTA (disodium salt)/0.05M-potassium phosphate, pH7.1; (c) sonication in 0.05M-potassium phosphate, pH7.1;(d)sonication in aq. Triton X-100 (0.1%). The membranes were pelleted by centrifugation at 100 000g for 15 min, between each step. Agglutination activity in the extracts was investigated by using trypsin-treated rabbit erythrocytes. 3. The addition of lactose to inner mitochondrial membrane resulted in the solubilization of part of the lectin activity, indicating that the protein was attached to the membrane via its carbohydrate-binding site. Pretreatment of the membranes with lactose before tha usual extraction procedure showed that lactose could extract lectins that normally required more harsh treatment of the membrane for solubilization. 4. Lectins extracted from inner membranes were purified by affinity chromatography on agarose gel. Polyacrylamide-gel electrophoresis of purified samples in sodium dodecyl sulphate indicated that at least part of the lectin present in inner mitochondrial membrane was identical with the R. communis agglutinin of mol.wt. 120 000. Images PLATE 1 PLATE 2 PMID:1008861

  7. Annotation and genetic diversity of the chicken collagenous lectins.

    PubMed

    Hamzić, Edin; Pinard-van der Laan, Marie-Hélène; Bed'Hom, Bertrand; Juul-Madsen, Helle Risdahl

    2015-06-01

    Collectins and ficolins are multimeric proteins present in various tissues and are actively involved in innate immune responses. In chickens, six different collagenous lectins have been characterized so far: mannose-binding lectin (MBL), surfactant protein A (SP-A), collectin 10 (COLEC10), collectin 11 (COLEC11), collectin 12 (COLEC12), lung lectin (LL) and one ficolin (FCN). However, the structural and functional features of the chicken collectins and ficolin are still not fully understood. Therefore, the aims of this study were: (i) to make an overview of the genetic structure and function of chicken collectins and the ficolin, (ii) to investigate the variation in the chicken collectins and the ficolin gene in different chicken populations, and (iii) to assess the presence of MBL gene variants in different chicken populations. We performed comparative genomic analysis using publically available data. The obtained results showed that collectins and ficolins have conserved protein sequences and gene structure across all vertebrate groups and this is especially notable for COLEC10, COLEC11 and COLEC12. For the purpose of studying the genetic variation, 179 animals from 14 populations were genotyped using 31 SNPs covering five genomic regions. The obtained results revealed low level of heterozygosity in the collagenous lectins except for the COLEC12 gene and the LL-SPA-MBL region compared to heterozygosity at neutral microsatellite markers. In addition, the MBL gene variants were assessed in different chicken populations based on the polymorphisms in the promoter region. We observed 10 previously identified MBL variants with A2/A8 and A4 as the most frequent alleles.

  8. Lectins as membrane components of mitochondria from Ricinus communis.

    PubMed

    Bowles, D J; Schnarrenberger, C; Kauss, H

    1976-11-15

    1. Mitochondria were isolated from developing endosperm of Ricinus communis and were fractionated into outer membrane and inner membrane. The relative purity of the two membrane fractions was determined by marker enzymes. The fractions were also examined by negative-stain electron microscopy. 2. Membrane fractions were sequentially extracted in the following way. (a) Suspension in 0.5M-potassium phosphate, pH7.1; (b)suspension in 0.1M-EDTA (disodium salt)/0.05M-potassium phosphate, pH7.1; (c) sonication in 0.05M-potassium phosphate, pH7.1;(d)sonication in aq. Triton X-100 (0.1%). The membranes were pelleted by centrifugation at 100 000g for 15 min, between each step. Agglutination activity in the extracts was investigated by using trypsin-treated rabbit erythrocytes. 3. The addition of lactose to inner mitochondrial membrane resulted in the solubilization of part of the lectin activity, indicating that the protein was attached to the membrane via its carbohydrate-binding site. Pretreatment of the membranes with lactose before tha usual extraction procedure showed that lactose could extract lectins that normally required more harsh treatment of the membrane for solubilization. 4. Lectins extracted from inner membranes were purified by affinity chromatography on agarose gel. Polyacrylamide-gel electrophoresis of purified samples in sodium dodecyl sulphate indicated that at least part of the lectin present in inner mitochondrial membrane was identical with the R. communis agglutinin of mol.wt. 120 000.

  9. A simple fibril and lectin model for cyst walls of Entamoeba and perhaps Giardia

    PubMed Central

    Samuelson, John; Robbins, Phillips

    2010-01-01

    Cyst walls of Entamoeba and Giardia protect them from environmental insults, stomach acids, and intestinal proteases. Each cyst wall contains a sugar homopolymer: chitin in Entamoeba and a unique N-acetylgalactosamine (GalNAc) homopolymer in Giardia. Entamoeba cyst wall proteins include Jacob lectins (carbohydrate-binding proteins) that cross-link chitin, chitinases that degrade chitin, and Jessie lectins that make walls impermeable. Giardia cyst wall proteins are also lectins that bind fibrils of the GalNAc homopolymer. While many of the details remain to be determined for the Giardia cyst wall, current data suggests a relatively simple fibril and lectin model for the Entamoeba cyst wall. PMID:20934911

  10. Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses.

    PubMed

    Opitz, Lars; Salaklang, Jatuporn; Büttner, Hermann; Reichl, Udo; Wolff, Michael W

    2007-01-15

    The presented study aims on the development of a capture step for the purification of cell culture derived influenza viruses using lectin affinity chromatography. Human influenza A/Puerto Rico/8/34 virus produced in Madin Darby canine kidney cells have been chosen as a model. The influenza A virus envelop possesses two viral glycoproteins: hemagglutinin and neuraminidase. Oligosaccharides of theses glycoproteins can be targeted as affinity ligands using specific lectins. First, lectins have been screened via lectin blots and spin columns. Adequate lectins have been chosen based on published glycan structures of hemagglutinin. The most specific binding was achieved via the galactose specific Erythrina cristagalli and Euonymus europaeus lectins. Second, the chromatographic separations characteristics of these lectins have been further determined via FPLC. These experiments revealed that the rate of hemagglutinin glycan binding to the ligands was higher with the E. europaeus compared to the E. cristagalli lectin. Third, viral recoveries in addition to the total protein and host cell DNA have been balanced in a series of E. europaeus lectin chromatography runs. The total protein and dsDNA content in the product fraction of the affinity chromatography was reduced from the starting conditions to 21% and 0.1%, respectively. The average viral recovery in the product fraction was 97%. SDS-PAGE analysis indicated that the majority of the eluted proteins were of viral origin. The reproducibility and column stability was confirmed in up to 25 runs applying six different virus product batches.

  11. The three-dimensional structure of codakine and related marine C-type lectins.

    PubMed

    Gourdine, Jean-Philippe; Markiv, Anatoly; Smith-Ravin, Juliette

    2007-10-01

    Codakine is a new Ca(2+)-dependent mannose-binding C-type lectin (MBL) isolated from the gill tissue of the tropical clam, Codakia orbicularis. Bioinformatic analyses with the BLAST program have revealed similarities with marine lectins involved in immunity whose three-dimensional (3D) structures were unknown up until recently. In this article, we present bioinformatic analyses of marine lectins that are homologous to codakine, in particular lectins from the sea worm Laxus oneistus, named mermaid. These lectins are involved in the symbiotic association with sulphur-oxidizing bacteria which are closely related to the C. orbicularis gill symbiont. Using homology modelling, folding that is characteristic of C-type lectins was observed in all the marine Ca(2+)-dependent lectins studied, with conservation of random coiled structures of the carbohydrate recognition domain (CRD) and Ca(2+)-binding sites. Like codakine, the marine lectins analysed contain a signal peptide commonly found in secreted and transmembrane proteins. The majority of the predictive 3D models established from the lectins exhibit a common feature, namely the involvement in invertebrate and vertebrate immunity (dendritic cell receptor, macrophage receptor, etc.). These bioinformatic analyses and the literature data support the hypothesis that codakine, like the L. oneistus mermaids, is probably involved in the cellular mediation of symbiosis and defence against pathogenic microorganisms.

  12. Parkia pendula seed lectin: potential use to treat cutaneous wounds in healthy and immunocompromised mice.

    PubMed

    Coriolano, Marília Cavalcanti; de Melo, Cristiane Moutinho Lagos; Silva, Flávio de Oliveira; Schirato, Giuliana Viegas; Porto, Camila Souza; dos Santos, Paulo Jorge Parreira; Correia, Maria Tereza dos Santos; Porto, Ana Lúcia Figueiredo; Carneiro-Leão, Ana Maria dos Anjos; Coelho, Luana Cassandra Breitenbach Barroso

    2014-03-01

    Parkia pendula seed lectin was used to treat cutaneous wounds of normal and immunocompromised mice, inducing cicatrization. Methotrexate (0.8 mg/kg/week) was used as immunosuppressive drug. Wounds were produced in the dorsal region (1 cm(2)) of female albino Swiss mice (Mus musculus), health and immunocompromised. Wounds were daily topically treated with 100 μL of the following solutions: (1) control (NaCl 0.15 M), (2) control Im (0.15 M NaCl), (3) P. pendula seed lectin (100 μg/mL), and (4) P. pendula seed lectin Im (100 μg/mL). Clinical evaluation was performed during 12 days. Biopsies for histopathology analysis and microbiological examinations were carried out in the second, seventh, and 12th days. The presence of edema and hyperemia was observed in all groups during inflammatory period. The first crust was detected from the second day, only in the groups treated with P. pendula seed lectin. Microbiological analysis of wounds from day 0 to day 2 did not show bacterium at P. pendula seed lectin group; however, Staphylococcus sp. was detected every day in the other groups. The lectin markedly induced a total wound closing at P. pendula seed lectin and P. pendula seed lectin Im groups on 11th day of evolution. The present study suggests that P. pendula seed lectin is a biomaterial potential to show pharmacological effect in the repair process of cutaneous wounds.

  13. Toxicity and binding profile of lectins from the Genus canavalia on brine shrimp.

    PubMed

    Arruda, Francisco Vassiliepe Sousa; Melo, Arthur Alves; Vasconcelos, Mayron Alves; Carneiro, Romulo Farias; Barroso-Neto, Ito Liberato; Silva, Suzete Roberta; Pereira-Junior, Francisco Nascimento; Nagano, Celso Shiniti; Nascimento, Kyria Santiago; Teixeira, Edson Holanda; Saker-Sampaio, Silvana; Sousa Cavada, Benildo; Sampaio, Alexandre Holanda

    2013-01-01

    Lectins are sugar-binding proteins widely distributed in nature with many biological functions. Although many lectins have a remarkable biotechnological potential, some of them can be cytotoxic. Thus, the aim of this study was to assess the toxicity of five lectins, purified from seeds of different species of Canavalia genus. In order to determine the toxicity, assays with Artemia nauplii were performed. In addition, a fluorescence assay was carried out to evaluate the binding of lectins to Artemia nauplii. In order to verify the relationship between the structure of lectins and their cytotoxic effect, structural analysis was carried out to evaluate the volume of the carbohydrate recognition domain (CRD) of each lectin. The results showed that all lectins exhibited different toxicities and bound to a similar area in the digestive tract of Artemia nauplii. Concerning the structural analysis, differences in spatial arrangement and volume of CRD may explain the variation of the toxicity exhibited by each lectin. To this date, this is the first study that establishes a link between toxicity and structure of CRD from Diocleinae lectins.

  14. Characterization of a new lectin involved in the protoplast regeneration of Bryopsis hypnoides

    NASA Astrophysics Data System (ADS)

    Niu, Jianfeng; Wang, Guangce; Lü, Fang; Zhou, Baicheng; Peng, Guang

    2009-09-01

    A group of coenocytic marine algae differs from higher plants, whose totipotency depends on an intact cell (or protoplast). Instead, this alga is able to aggregate its extruded protoplasm in sea water and generate new mature individuals. It is thought that lectins play a key role in the aggregation process. We purified a lectin associated with the aggregation of cell organelles in Bryopsis hypnoides. The lectin was ca. 27 kDa with a pI between pH 5 and pH 6. The absence of carbohydrate suggested that the lectin was not a glycoprotein. The hemagglutinating activity (HA) of the lectin was not dependent on the presence of divalent cations and was inhibited by N-Acetylgalactosamine, N-Acetylglucosamine, and the glycoprotein bovine submaxillary mucin. The lectin preferentially agglutinated Gram-negative bacterium. The HA of this lectin was stable between pH 4 to pH 10. Cell organelles outside the cytoplasm were agglutinated by the addition of lectin solution (0.5 mg ml-1). Our results suggest that the regeneration of B. hypnoides is mediated by this lectin. We also demonstrated that the formation of cell organelle aggregates was inhibited by nigericin in natural seawater (pH 8.0). Given that nigericin dissipates proton gradients across the membrane, we hypothesize that the aggregation of cell organelles was proton-gradient dependent.

  15. Preparation of affinity adsorbents and purification of lectins from natural sources.

    PubMed

    Date, Kimie; Ogawa, Haruko

    2014-01-01

    Lectins are purified by affinity chromatography to take advantage of their carbohydrate-specific interactions. Highly efficient affinity adsorbents are powerful tools to obtain homogeneous lectins with distinct specificities. Here, we describe three methods to prepare affinity adsorbents by immobilizing carbohydrates or glycoconjugates on agarose gel beads. Because the ligands are immobilized via a stable and nonionic linkage under mild conditions, the adsorbents possess high binding capacity for lectins with low nonspecific adsorption and can withstand repeated use. The procedures require neither specialized techniques and apparatus nor highly toxic compounds. Using these adsorbents, many plant and animal lectins can be purified in a few steps.

  16. Cloning and characterization of the lectin cDNA clones from onion, shallot and leek.

    PubMed

    Van Damme, E J; Smeets, K; Engelborghs, I; Aelbers, H; Balzarini, J; Pusztai, A; van Leuven, F; Goldstein, I J; Peumans, W J

    1993-10-01

    Characterization of the lectins from onion (Allium cepa), shallot (A. ascalonicum) and leek (A. porrum) has shown that these lectins differ from previously isolated Alliaceae lectins not only in their molecular structure but also in their ability to inhibit retrovirus infection of target cells. cDNA libraries constructed from poly(A)-rich RNA isolated from young shoots of onion, shallot and leek were screened for lectin cDNA clones using colony hybridization. Sequence analysis of the lectin cDNA clones from these three species revealed a high degree of sequence similarity both at the nucleotide and at the amino acid level. Apparently the onion, shallot and leek lectins are translated from mRNAs of ca. 800 nucleotides. The primary translation products are preproproteins (ca. 19 kDa) which are converted into the mature lectin polypeptides (12.5-13 kDa) after post-translational modifications. Southern blot analysis of genomic DNA has shown that the lectins are most probably encoded by a family of closely related genes which is in good agreement with the sequence heterogeneity found between different lectin cDNA clones of one species.

  17. Responsive Photonic Crystal Carbohydrate Hydrogel Sensor Materials for Selective and Sensitive Lectin Protein Detection.

    PubMed

    Cai, Zhongyu; Sasmal, Aniruddha; Liu, Xinyu; Asher, Sanford A

    2017-10-04

    Lectin proteins, such as the highly toxic lectin protein, ricin, and the immunochemically important lectin, jacalin, play significant roles in many biological functions. It is highly desirable to develop a simple but efficient method to selectively detect lectin proteins. Here we report the development of carbohydrate containing responsive hydrogel sensing materials for the selective detection of lectin proteins. The copolymerization of a vinyl linked carbohydrate monomer with acrylamide and acrylic acid forms a carbohydrate hydrogel that shows specific "multivalent" binding to lectin proteins. The resulting carbohydrate hydrogels are attached to 2-D photonic crystals (PCs) that brightly diffract visible light. This diffraction provides an optical readout that sensitively monitors the hydrogel volume. We utilize lactose, galactose, and mannose containing hydrogels to fabricate a series of 2-D PC sensors that show strong selective binding to the lectin proteins ricin, jacalin, and concanavalin A (Con A). This binding causes a carbohydrate hydrogel shrinkage which significantly shifts the diffraction wavelength. The resulting 2-D PC sensors can selectively detect the lectin proteins ricin, jacalin, and Con A. These unoptimized 2-D PC hydrogel sensors show a limit of detection (LoD) of 7.5 × 10(-8) M for ricin, a LoD of 2.3 × 10(-7) M for jacalin, and a LoD of 3.8 × 10(-8) M for Con A, respectively. This sensor fabrication approach may enable numerous sensors for the selective detection of numerous lectin proteins.

  18. Toxicity and Binding Profile of Lectins from the Genus Canavalia on Brine Shrimp

    PubMed Central

    Arruda, Francisco Vassiliepe Sousa; Melo, Arthur Alves; Vasconcelos, Mayron Alves; Carneiro, Romulo Farias; Barroso-Neto, Ito Liberato; Silva, Suzete Roberta; Pereira-Junior, Francisco Nascimento; Nagano, Celso Shiniti; Nascimento, Kyria Santiago; Teixeira, Edson Holanda; Saker-Sampaio, Silvana; Sousa Cavada, Benildo; Sampaio, Alexandre Holanda

    2013-01-01

    Lectins are sugar-binding proteins widely distributed in nature with many biological functions. Although many lectins have a remarkable biotechnological potential, some of them can be cytotoxic. Thus, the aim of this study was to assess the toxicity of five lectins, purified from seeds of different species of Canavalia genus. In order to determine the toxicity, assays with Artemia nauplii were performed. In addition, a fluorescence assay was carried out to evaluate the binding of lectins to Artemia nauplii. In order to verify the relationship between the structure of lectins and their cytotoxic effect, structural analysis was carried out to evaluate the volume of the carbohydrate recognition domain (CRD) of each lectin. The results showed that all lectins exhibited different toxicities and bound to a similar area in the digestive tract of Artemia nauplii. Concerning the structural analysis, differences in spatial arrangement and volume of CRD may explain the variation of the toxicity exhibited by each lectin. To this date, this is the first study that establishes a link between toxicity and structure of CRD from Diocleinae lectins. PMID:24380079

  19. Microencapsulation of lectin anti-cancer agent and controlled release by alginate beads, biosafety approach.

    PubMed

    El-Aassar, M R; Hafez, Elsayed E; El-Deeb, Nehal M; Fouda, Moustafa M G

    2014-08-01

    Hepatocellular carcinoma (HCC) is considered as one of the most aggressive cancer worldwide. In Egypt, the prevalence of HCC is increasing during last years. Recently, drug-loaded microparticles were used to improve the efficiency of various medical treatments. This study is designed to evaluate the anticancer potentialities of lectins against HCC while hinting to its safety usage. The aim is also extended to encapsulate lectins in alginate microbeads for oral drug delivery purposes. The extracted lectins showed anti-proliferative effect against HCC with a percentage of 60.76% by using its nontoxic dose with an up-regulation of P53 gene expression. Concerning the handling of lectin alginate microbeads for oral drug delivery, the prepared lectin alginate beads were ∼100μm in diameter. The efficiency of the microcapsules was checked by scanning electron microscopy, the SEM showed the change on the alginate beads surface revealing the successful lectin encapsulation. The release of lectins from the microbeads depended on a variety of factors as the microbeads forming carriers and the amount-encapsulated lectins. The Pisum sativum extracted lectins may be considered as a promising agent in controlling HCC and this solid dosage form could be suitable for oral administration complemented with/or without the standard HCC drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Purification, characterization, and mitogenic potential of a mucin-specific mycelial lectin from Aspergillus sparsus.

    PubMed

    Singh, Ram Sarup; Bhari, Ranjeeta; Kaur, Rumeet

    2015-02-01

    Lectins are carbohydrate binding proteins or glycoproteins that bind reversibly to specific carbohydrates present on the apposing cells, which is responsible for their ability to agglutinate red blood cells, lymphocytes, fibroblasts, etc. Due to their carbohydrate specificity, lectins have been used for purification and characterization of glycoproteins like antibodies, cytokines, tumor-associated glycoproteins, hormones, inhibitors, growth factors, various enzymes, membrane proteins (receptors), or even toxins and viruses. In the present study, a mycelial lectin from Aspergillus sparsus was purified, characterized, and evaluated for its mitogenic potential. Lectin could be effectively purified 17.8-fold in a single-step using affinity chromatography on mucin-sepharose column. Lectin migrated as a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with an apparent molecular mass of 100.2 kDa. Lectin showed pH optima of 6.5-8.0, and optimum temperature was determined to be 20-30 °C. Lectin was stable within a pH range of 5.5-10.0 and showed fairly good thermostability. Lectin activity was unaffected in the presence of EDTA, while activity reduced upon interaction with denaturants. MTT assay revealed strong mitogenic potential of A. sparsus lectin at a concentration up to 100 μg/ml.

  1. Bitter-sweet symphony: glycan-lectin interactions in virus biology.

    PubMed

    Van Breedam, Wander; Pöhlmann, Stefan; Favoreel, Herman W; de Groot, Raoul J; Nauwynck, Hans J

    2014-07-01

    Glycans are carbohydrate modifications typically found on proteins or lipids, and can act as ligands for glycan-binding proteins called lectins. Glycans and lectins play crucial roles in the function of cells and organs, and in the immune system of animals and humans. Viral pathogens use glycans and lectins that are encoded by their own or the host genome for their replication and spread. Recent advances in glycobiological research indicate that glycans and lectins mediate key interactions at the virus-host interface, controlling viral spread and/or activation of the immune system. This review reflects on glycan-lectin interactions in the context of viral infection and antiviral immunity. A short introduction illustrates the nature of glycans and lectins, and conveys the basic principles of their interactions. Subsequently, examples are discussed highlighting specific glycan-lectin interactions and how they affect the progress of viral infections, either benefiting the host or the virus. Moreover, glycan and lectin variability and their potential biological consequences are discussed. Finally, the review outlines how recent advances in the glycan-lectin field might be transformed into promising new approaches to antiviral therapy. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  2. Functional recombinants designed from a fetuin/asialofetuin-specific marine algal lectin, rhodobindin.

    PubMed

    Han, Jong Won; Jung, Min Gui; Shim, Eun Young; Shim, Jun Bo; Kim, Young Min; Kim, Gwang Hoon

    2015-04-13

    Plant lectins have attracted much attention for biomedical applications including targeted drug delivery system and therapy against tumors and microbial infections. The main problem of using lectins as a biomedical tool is a batch-to-batch variation in isoforms content. The production of lectins using recombination tools has the advantage of obtaining high amounts of proteins with more precise properties, but there are only a handful of functional recombinant lectins presently available. A fetuin/asialo-fetuin specific lectin, Rhodobindin, has unique tandem repeats structure which makes it useful in exploiting for recombinant lectin. We developed three functional recombinant lectins using E. coli expression system: one from full cDNA sequence and two from fragmentary sequences of Rhodobindin. Hemagglutinating activity and solubility of the recombinant lectins were highest at OD 0.7 cell concentration at 20 °C. The optimized process developed in this study was suitable for the quality-controlled production of high amounts of soluble recombinant lectins.

  3. Functional Recombinants Designed from a Fetuin/Asialofetuin-Specific Marine Algal Lectin, Rhodobindin

    PubMed Central

    Han, Jong Won; Jung, Min Gui; Shim, Eun Young; Shim, Jun Bo; Kim, Young Min; Kim, Gwang Hoon

    2015-01-01

    Plant lectins have attracted much attention for biomedical applications including targeted drug delivery system and therapy against tumors and microbial infections. The main problem of using lectins as a biomedical tool is a batch-to-batch variation in isoforms content. The production of lectins using recombination tools has the advantage of obtaining high amounts of proteins with more precise properties, but there are only a handful of functional recombinant lectins presently available. A fetuin/asialo-fetuin specific lectin, Rhodobindin, has unique tandem repeats structure which makes it useful in exploiting for recombinant lectin. We developed three functional recombinant lectins using E. coli expression system: one from full cDNA sequence and two from fragmentary sequences of Rhodobindin. Hemagglutinating activity and solubility of the recombinant lectins were highest at OD 0.7 cell concentration at 20 °C. The optimized process developed in this study was suitable for the quality-controlled production of high amounts of soluble recombinant lectins. PMID:25871294

  4. Detection, purification and characterization of a lectin from freshwater green algae Spirogyra spp.

    PubMed

    Oliveira, Antônia S DE; Lóssio, Cláudia F; Rangel, Anne J; Martins, Maria G Q; Nascimento, Fernando E P DO; Andrade, Maria L L DE; Cavada, Benildo S; Lacerda, Sírleis R; Nascimento, Kyria S DO

    2017-08-31

    Freshwater algae are rich sources of structurally biologically active metabolites, such as fatty acids, steroids, carotenoids and polysaccharides. Among these metabolites, lectins stand out. Lectins are proteins or glycoproteins of non-immune origin which bind to carbohydrates or glycoconjugates, without changing ligand structure. Many studies have reported on the use of Spirogyra spp. as effective bioindicators of heavy metals; however, reports on Spirogyra molecular bioprospecting are quite limited. Therefore, this study aimed to detect, isolate, purify and characterize a lectin present in the freshwater green algae Spirogyra. Presence of the lectin protein in the extract was detected by hemagglutination assays. Subsequently, the protein extract was subjected to a sugar inhibition assay to identify the lectin-specific carbohydrate. Following this, the extract was applied to a guar gum column to afford the pure lectin. The lectin was inhibited by N-acetyl-glucosamine and N-acetyl-beta-D-mannose, but more strongly by D-galactose. The apparent molecular mass of the purified lectin was evaluated by Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Electrophoretic analysis revealed a single protein band with an apparent molecular mass of 56 kDa. Thus, it could be concluded that a lectin was purified from Spirogyra spp.

  5. A lectin-mediated resistance of higher fungi against predators and parasites.

    PubMed

    Bleuler-Martínez, S; Butschi, A; Garbani, M; Wälti, M A; Wohlschlager, T; Potthoff, E; Sabotiĉ, J; Pohleven, J; Lüthy, P; Hengartner, M O; Aebi, M; Künzler, M

    2011-07-01

    Fruiting body lectins are ubiquitous in higher fungi and characterized by being synthesized in the cytoplasm and up-regulated during sexual development. The function of these lectins is unclear. A lack of phenotype in sexual development upon inactivation of the respective genes argues against a function in this process. We tested a series of characterized fruiting body lectins from different fungi for toxicity towards the nematode Caenorhabditis elegans, the mosquito Aedes aegypti and the amoeba Acanthamoeba castellanii. Most of the fungal lectins were found to be toxic towards at least one of the three target organisms. By altering either the fungal lectin or the glycans of the target organisms, or by including soluble carbohydrate ligands as competitors, we demonstrate that the observed toxicity is dependent on the interaction between the fungal lectins and specific glycans in the target organisms. The toxicity was found to be dose-dependent such that low levels of lectin were no longer toxic but still led to food avoidance by C. elegans. Finally, we show, in an ecologically more relevant scenario, that challenging the vegetative mycelium of Coprinopsis cinerea with the fungal-feeding nematode Aphelenchus avenae induces the expression of the nematotoxic fruiting body lectins CGL1 and CGL2. Based on these findings, we propose that filamentous fungi possess an inducible resistance against predators and parasites mediated by lectins that are specific for glycans of these antagonists.

  6. Comparative biochemical analysis of lectin and nuclease from Chelidonium majus L.

    PubMed

    Fik, E; Dalgalarrondo, M; Haertlé, T; Goździcka-Józefiak, A

    2000-01-01

    It has been recently recognized that lectins exhibit other activities besides hemagglutination. Previously we have found that purified lectin from Chelidonium majus showed DNase activity (Fik, Goździcka-Józefiak & Kedzia, 1995, Herba Polon. 41, 84-95). Comparison of lectin and DNase from the sap from leaves and roots of Chelidonium majus proved that both these compounds are composed of 24 kDa monomer subunits which have an identical N-terminal sequence but differ in amino-acid composition and degree of glycosylation. Possible interrelationship between lectin and DNase is discussed.

  7. Ganoderma lucidum: a source for novel bioactive lectin.

    PubMed

    U Girjal, Vinay; Neelagund, Shivayogeeswar; Krishnappa, Madappa

    2011-11-01

    Ganoderma lucidum is known for its high medicinal value, clinically used in treatment for various diseases. We have selected this mushroom for isolation of novel bioactive lectin. The isolation procedure comprised of ion-exchange chromatography on DEAE- cellulose and affinity chromatography on Affi-gel blue gel. Purified lectin was monomer with a molecular mass of 15 kDa, determined by SDS-PAGE, Gel filtration, MALDI-ToF. It showed hemagglutinating activity against both human and animal erythrocytes. The hemagglutination activity was not inhibited by simple sugars but inhibited by glycoproteins. The activity was maximal at pH range 4.0-9.0 and at temperature up to 60° C. The hemagglutination activity was stable even in the presence of 10mM EDTA and other divalent metal cations such as CaCl2, MgCl2, ZnCl2, and MnCl2. Lectin was shown antifungal activity against following pathogens Fusarium oxysporium, Penicillium chrysogenum, Aspergillus Niger, Colletotrichum musae, Botrytis cinerea, Trichophyton rubrum, Trichophyton tonsurans, Trichophyton interdigitale, Epidermophyton floccosum and Microsporum canis.

  8. Regional differences in lectin binding patterns of vestibular hair cells

    NASA Technical Reports Server (NTRS)

    Baird, Richard A.; Schuff, N. R.; Bancroft, J.

    1994-01-01

    Surface glycoconjugates of hair cells and supporting cells in the vestibular endorgans of the bullfrog were identified using biotinylated lectins with different carbohydrate specificities. Lectin binding in hair cells was consistent with the presence of glucose and mannose (CON A), galactose (RCA-I), N-acetylgalactosamine (VVA), but not fucose (UEA-I) residues. Hair cells in the bullfrog sacculus, unlike those in the utriculus and semicircular canals, did not stain for N-acetylglucosamine (WGA) or N-acetylgalactosamine (VVA). By contrast, WGA and, to a lesser extent, VVA, differentially stained utricular and semicircular canal hair cells, labeling hair cells located in peripheral, but not central, regions. In mammals, WGA uniformly labeled Type 1 hair cells while labeling, as in the bullfrog, Type 2 hair cells only in peripheral regions. These regional variations were retained after enzymatic digestion. We conclude that vestibular hair cells differ in their surface glycoconjugates and that differences in lectin binding patterns can be used to identify hair cell types and to infer the epithelial origin of isolated vestibular hair cells.

  9. Regional differences in lectin binding patterns of vestibular hair cells

    NASA Technical Reports Server (NTRS)

    Baird, R. A.; Schuff, N. R.; Bancroft, J.

    1993-01-01

    Surface glycoconjugates of hair cells and supporting cells in the vestibular endorgans of the bullfrog were identified using biotinylated lectins with different carbohydrate specificities. Lectin binding in hair cells was consistent with the presence of glucose and mannose (CON A), galactose (RCA-I), N-acetylglucosamine (WGA), N-acetylgalactosamine (VVA), but not fucose (UEA-I) residues. Hair cells in the bullfrog sacculus, unlike those in the utriculus and semicircular canals, did not strain for N-acetylglucosamine (WGA) or N-acetylgalactosamine (VVA). By contrast, WGA and, to a lesser extent, VVA, differentially stained utricular and semicircular canal hair cells, labeling hair cells located in peripheral, but not central, regions. In mammals, WGA uniformly labeled Type I hair cells while labeling, as in the bullfrog, Type II hair cells only in peripheral regions. These regional variations were retained after enzymatic digestion. We conclude that vestibular hair cells differ in their surface glycoconjugates and that differences in lectin binding patterns can be used to identify hair cell types and to infer the epithelial origin of isolated vestibular hair cells.

  10. Antibacterial activity of a lectin-like Burkholderia cenocepacia protein

    PubMed Central

    Ghequire, Maarten G K; Canck, Evelien; Wattiau, Pierre; Winge, Iris; Loris, Remy; Coenye, Tom; Mot, René

    2013-01-01

    Abstract Bacteriocins of the LlpA family have previously been characterized in the γ-proteobacteria Pseudomonas and Xanthomonas. These proteins are composed of two MMBL (monocot mannose-binding lectin) domains, a module predominantly and abundantly found in lectins from monocot plants. Genes encoding four different types of LlpA-like proteins were identified in genomes from strains belonging to the Burkholderia cepacia complex (Bcc) and the Burkholderia pseudomallei group. A selected recombinant LlpA-like protein from the human isolate Burkholderia cenocepacia AU1054 displayed narrow-spectrum genus-specific antibacterial activity, thus representing the first functionally characterized bacteriocin within this β-proteobacterial genus. Strain-specific killing was confined to other members of the Bcc, with mostly Burkholderia ambifaria strains being susceptible. In addition to killing planktonic cells, this bacteriocin also acted as an antibiofilm agent. Bacteriocins mediate highly selective antagonism among closely related bacteria but such antimicrobial proteins have not yet been reported in Burkholderia. We identified a lectin-like protein of the LlpA family in a Burkholderia cenocepacia human isolate that strain-specifically and selectively kills planktonic and biofilm cells of other Burkholderia cepacia complex members. PMID:23737242

  11. Labeling of lectin receptors during the cell cycle.

    PubMed

    Garrido, J

    1976-12-01

    Labeling of lectin receptors during the cell cycle. (Localizabión de receptores para lectinas durante el ciclo celular). Arch. Biol. Med. Exper. 10: 100-104, 1976. The topographic distribution of specific cell surface receptors for concanavalin A and wheat germ agglutinin was studied by ultrastructural labeling in the course of the cell cycle. C12TSV5 cells were synchronized by double thymidine block or mechanical selection (shakeoff). They were labeled by means of lectin-peroxidase techniques while in G1 S, G2 and M phases of the cycle. The results obtained were similar for both lectins employed. Interphase cells (G1 S, G2) present a stlihtly discontinous labeling pattern that is similar to the one observed on unsynchronized cells of the same line. Cells in mitosis, on the contrary, present a highly discontinous distribution of reaction product. This pattern disappears after the cells enters G1 and is not present on mitotic cells fixed in aldehyde prior to labeling.

  12. Regional differences in lectin binding patterns of vestibular hair cells

    NASA Technical Reports Server (NTRS)

    Baird, R. A.; Schuff, N. R.; Bancroft, J.

    1993-01-01

    Surface glycoconjugates of hair cells and supporting cells in the vestibular endorgans of the bullfrog were identified using biotinylated lectins with different carbohydrate specificities. Lectin binding in hair cells was consistent with the presence of glucose and mannose (CON A), galactose (RCA-I), N-acetylglucosamine (WGA), N-acetylgalactosamine (VVA), but not fucose (UEA-I) residues. Hair cells in the bullfrog sacculus, unlike those in the utriculus and semicircular canals, did not strain for N-acetylglucosamine (WGA) or N-acetylgalactosamine (VVA). By contrast, WGA and, to a lesser extent, VVA, differentially stained utricular and semicircular canal hair cells, labeling hair cells located in peripheral, but not central, regions. In mammals, WGA uniformly labeled Type I hair cells while labeling, as in the bullfrog, Type II hair cells only in peripheral regions. These regional variations were retained after enzymatic digestion. We conclude that vestibular hair cells differ in their surface glycoconjugates and that differences in lectin binding patterns can be used to identify hair cell types and to infer the epithelial origin of isolated vestibular hair cells.

  13. Fluorescent lectins for local in vivo visualization of peripheral nerves.

    PubMed

    KleinJan, Gijs Hendrik; Buckle, Tessa; van Willigen, Danny Michel; van Oosterom, Matthias Nathanaël; Spa, Silvia Johara; Kloosterboer, Harmen Egbert; van Leeuwen, Fijs Willem Bernhard

    2014-07-08

    Damage to peripheral nerves caused during a surgical intervention often results in function loss. Fluorescence imaging has the potential to improve intraoperative identification and preservation of these structures. However, only very few nerve targeting agents are available. This study describes the in vivo nerve staining capabilities of locally administered fluorescent lectin-analogues. To this end WGA, PNA, PHA-L and LEL were functionalized with Cy5 (λex max 640 nm; λem max 680 nm). Transfer of these imaging agents along the sciatic nerve was evaluated in Thy1-YFP mice (n = 12) after intramuscular injection. Migration from the injection site was assessed in vivo using a laboratory fluorescence scanner and ex vivo via fluorescence confocal microscopy. All four lectins showed retrograde movement and staining of the epineurium with a signal-to-muscle ratio of around two. On average, the longest transfer distance was obtained with WGA-Cy5 (0.95 cm). Since WGA also gave minimal uptake in the lymphatic system, this lectin type revealed the highest potential as a migration imaging agent to visualize nerves.

  14. Engineering a Therapeutic Lectin by Uncoupling Mitogenicity from Antiviral Activity

    PubMed Central

    Swanson, Michael D.; Boudreaux, Daniel M.; Salmon, Loïc; Chugh, Jeetender; Winter, Harry C.; Meagher, Jennifer L.; André, Sabine; Murphy, Paul V.; Oscarson, Stefan; Roy, René; King, Steven; Kaplan, Mark H.; Goldstein, Irwin J.; Tarbet, E. Bart; Hurst, Brett L.; Smee, Donald F.; de la Fuente, Cynthia; Hoffmann, Hans-Heinrich; Xue, Yi; Rice, Charles M.; Schols, Dominique; Garcia, J. Victor; Stuckey, Jeanne A.; Gabius, Hans-Joachim; Al-Hashimi, Hashim M.; Markovitz, David M.

    2015-01-01

    Summary A key effector route of the Sugar Code involves lectins that exert crucial regulatory controls by targeting distinct cellular glycans. We demonstrate that a single amino acid substitution in a banana lectin, replacing histidine 84 with a threonine, significantly reduces its mitogenicity while preserving its broad-spectrum antiviral potency. X-ray crystallography, NMR spectroscopy, and glycocluster assays reveal that loss of mitogenicity is strongly correlated with loss of pi-pi stacking between aromatic amino acids H84 and Y83, which removes a wall separating two carbohydrate binding sites, thus diminishing multivalent interactions. On the other hand, monovalent interactions and antiviral activity are preserved by retaining other wild-type conformational features and possibly through unique contacts involving the T84 side chain. Through such fine-tuning, target selection and downstream effects of a lectin can be modulated so as to knock down one activity while preserving another, thus providing tools for therapeutics and for understanding the Sugar Code. PMID:26496612

  15. Crystal structure of a symbiosis-related lectin from octocoral.

    PubMed

    Kita, Akiko; Jimbo, Mitsuru; Sakai, Ryuichi; Morimoto, Yukio; Miki, Kunio

    2015-09-01

    D-Galactose-binding lectin from the octocoral, Sinularia lochmodes (SLL-2), distributes densely on the cell surface of microalgae, Symbiodinium sp., an endosymbiotic dinoflagellate of the coral, and is also shown to be a chemical cue that transforms dinoflagellate into a non-motile (coccoid) symbiotic state. SLL-2 binds with high affinity to the Forssman antigen (N-acetylgalactosamine(GalNAc)α1-3GalNAcβ1-3Galα1-4Galβ1-4Glc-ceramide), and the presence of Forssman antigen-like sugar on the surface of Symbiodinium CS-156 cells was previously confirmed. Here we report the crystal structures of SLL-2 and its GalNAc complex as the first crystal structures of a lectin involved in the symbiosis between coral and dinoflagellate. N-Linked sugar chains and a galactose derivative binding site common to H-type lectins were observed in each monomer of the hexameric SLL-2 crystal structure. In addition, unique sugar-binding site-like regions were identified at the top and bottom of the hexameric SLL-2 structure. These structural features suggest a possible binding mode between SLL-2 and Forssman antigen-like pentasaccharide.

  16. The mannose-specific lectins from ramsons (Allium ursinum L.) are encoded by three sets of genes.

    PubMed

    Van Damme, J M; Smeets, K; Torrekens, S; Van Leuven, F; Peumans, W J

    1993-10-01

    Lectin cDNA clones encoding the two mannose-binding lectins from ramsons (allium ursinum L.) bulbs, AUAI and AUAII (AUA, Allium ursinum agglutinin), were isolated and characterized. Sequence comparison of the different cDNA clones isolated revealed three types of lectin clones called LECAUAG0, LECAUAG1 and LECAUAG2, which besides the obvious differences in their sequences also differ from each other in the number of potential glycosylation sites within the C-terminal peptide of the lectin precursor. In vivo biosynthesis studies of the ramson lectins have shown that glycosylated lectin precursors occur in the organelle fraction of radioactively labeled ramson bulbs. Despite the similarities between the A. ursinum and the A. sativum (garlic) lectins at the protein level, molecular cloning of the two ramson lectins has shown that the lectin genes in A. ursinum are organized differently. Whereas in A. sativum the lectin polypeptides of the heterodimeric ASAI are encoded by one large precursor, those of the heterodimeric AUAI lectin are derived from two different precursors. These results are confirmed by Northern blot hybridization of A. ursinum RNA which, after hybridization with a labeled lectin cDNA, reveals only one band of 800 nucleotides in contrast to A. sativum RNA which yields two bands of 1400 and 800 nucleotides. Furthermore it is shown that the two mannose-binding lectins are differentially expressed.

  17. Characterization of IgE-binding epitopes of peanut (Arachis hypogaea) PNA lectin allergen cross-reacting with other structurally related legume lectins.

    PubMed

    Rougé, Pierre; Culerrier, Raphaël; Granier, Claude; Rancé, Fabienne; Barre, Annick

    2010-08-01

    Sera from peanut allergic patients contain IgE that specifically interact with the peanut lectin PNA and other closely related legume lectins like LcA from lentil, PsA from pea and PHA from kidney bean. The IgE-binding activity of PNA and legume lectins was assessed by immunoblotting, surface plasmon resonance (SPR) and ELISA measurements, using sera from peanut allergic patients as a IgE source. This IgE-binding cross-reactivity most probably depends on the occurrence of structurally related epitopes that have been identified on the molecular surface of PNA and other legume lectins. These epitopes definitely differ from those responsible for the allergenicity of the major allergens Ara h 1, Ara h 2 and Ara h 3, also recognized by the IgE-containing sera of peanut allergic patients. Peanut lectin PNA and other legume lectins have been characterized as potential allergens for patients allergic to edible legume seeds. However, the clinical significance of the lectin-IgE interaction has to be addressed. (c) 2010 Elsevier Ltd. All rights reserved.

  18. Interaction of linear manno-oligosaccharides with three mannose-specific bulb lectins. Comparison with mannose/glucose-binding lectins.

    PubMed

    Kaku, H; Goldstein, I J

    1992-05-22

    Three new mannose-binding lectins, isolated from daffodil (NPA), amaryllis (HHA), and snowdrop (GNA) bulbs, are capable of precipitating with a linear mannopentaose (Man alpha 1-3Man alpha 1-3Man alpha 1-3Man alpha 1-2Man). NPA and HHA reacted strongly with the mannopentaose whereas GNA gave a precipitate only at concentrations greater than 500 microM. A phosphate group at C-6 of the nonreducing terminal mannosyl group prevented precipitation in all three cases. The reduced (NaBH4) mannopentaose, Man4Man-ol, did not precipitate with GNA or NPA, but was active with HHA. This activity was lost when Man4Man-ol was converted (NaIO4 then NaBH4; mild acid hydrolysis of the reduced product) into trisaccharide derivatives. With alpha-D-Manp-OMe the three lectins gave UV difference spectra having large positive peaks at 292-293 and 283-284 nm, and a small positive peak at 275 nm, characteristic of tryptophanyl and tyrosyl residues. The association constants for the interaction with alpha-D-Manp-OMe were very low (NPA, 86; HHA, 66; and GNA, 41 M-1), but the lectins bound methyl (1----3)-alpha-mannobioside with increased affinity (K for NPA 540, for HHA 2400, and for GNA 200 M-1). The bulb lectins lack binding sites for hydrophobic ligands, as judged by their failure to interact with the fluorescent probes 8-anilino-1-napthalenesulfonic acid (ANS) and 6-p-toluidino-2-naphthalenesulfonic acid (TNS).

  19. Identification of a Macrobrachium nipponense C-type lectin with a close evolutionary relationship to vertebrate lectins.

    PubMed

    Huang, Xin; Li, Tingting; Jin, Min; Yin, Shaowu; Wang, Wen; Ren, Qian

    2017-07-01

    C-type lectins (CTLs) are involved in the innate immune defense of vertebrates and invertebrates against invading pathogens. This study cloned and characterized a novel C-type lectin (MnCTL) of the oriental river prawn, Macrobrachium nipponense. The cloned MnCTL cDNA encompasses an open reading frame of 774 nucleotides and encodes polypeptides of 257 residues. The deduced MnCTL protein contains a single carbohydrate recognition domain (CRD) with an EPN (Glu-Pro-Asn) motif in calcium-binding site 2. Phylogenetic analysis indicated that MnCTL has a closer evolutionary relationship with vertebrate lectins than with invertebrate lectins. Tissue expression analysis showed that high levels of MnCTL are ubiquitously distributed in the gills and stomach of M. nipponense. Quantitative real-time RT-PCR (qRT-PCR) analysis showed that MnCTL expression was up-regulated by bacteria or white spot syndrome virus (WSSV) challenge. Knock-down of the MnCTL gene in WSSV-challenged prawns significantly decreased MnALF1 and MnALF2 transcript levels. The recombinant MnCRD (rMnCRD) agglutinated both Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Vibrio parahaemolyticus) in the presence of calcium. Furthermore, rMnCRD could bind to all the tested bacteria with different activities. The sugar-binding assay showed that rMnCRD was able to bind lipopolysaccharide and peptidoglycan in a concentration-dependent manner. In addition, rMnCRD could accelerate bacterial clearance. On the contrary, MnCTL silencing by dsRNA interference could weaken the bacterial clearance ability. All these findings implicated MnCTL were involved in the antiviral and antibacterial innate immunity of M. nipponense. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Caulobacters in the Marine Environment.

    DTIC Science & Technology

    1987-01-01

    LECIIN> PEANUT DOLICHOS SOYBEAN CONCONAV- ULEX WHEAT GERM RICINUS STRAIN AGGLUTININ BIFLORUS AGGLUTININ ALINA EUROPEAUS AGGLUTININ COMMUNI ~i MCS 3 MCS3...LECTIN> fPEANUT DOLICHOS SOYBEAN CONCONAV- ULEX WHEAT GERM RICINUS STRAI N lAGGLUTININ BIFLORUS AGGLUTININ ALINA EUROPEAUS1

  1. Near-planar Solution Structures of Mannose-binding Lectin Oligomers Provide Insight on Activation of Lectin Pathway of Complement

    PubMed Central

    Miller, Ami; Phillips, Anna; Gor, Jayesh; Wallis, Russell; Perkins, Stephen J.

    2012-01-01

    The complement system is a fundamental component of innate immunity that orchestrates complex immunological and inflammatory processes. Complement comprises over 30 proteins that eliminate invading microorganisms while maintaining host cell integrity. Protein-carbohydrate interactions play critical roles in both the activation and regulation of complement. Mannose-binding lectin (MBL) activates the lectin pathway of complement via the recognition of sugar arrays on pathogenic surfaces. To determine the solution structure of MBL, synchrotron x-ray scattering and analytical ultracentrifugation experiments showed that the carbohydrate-recognition domains in the MBL dimer, trimer, and tetramer are positioned close to each other in near-planar fan-like structures. These data were subjected to constrained modeling fits. A bent structure for the MBL monomer was identified starting from two crystal structures for its carbohydrate-recognition domain and its triple helical region. The MBL monomer structure was used to identify 10–12 near-planar solution structures for each of the MBL dimers, trimers, and tetramers starting from 900 to 6,859 randomized structures for each. These near-planar fan-like solution structures joined at an N-terminal hub clarified how the carbohydrate-recognition domain of MBL binds to pathogenic surfaces. They also provided insight on how MBL presents a structural template for the binding and auto-activation of the MBL-associated serine proteases to initiate the lectin pathway of complement activation. PMID:22167201

  2. Evidence for export of a muscle lectin from cytosol to extracellular matrix and for a novel secretory mechanism

    SciTech Connect

    Cooper, D.N.; Barondes, S.H. )

    1990-05-01

    A soluble lactose-binding lectin with subunit Mr of 14,500 is believed to function by interacting with extracellular glycoconjugates, because it has been detected extracellularly by immunohistochemistry. This localization has been questioned, however, since the lectin lacks a secretion signal sequence, which challenges the contention that it is secreted. We have demonstrated externalization of this lectin from C2 mouse muscle cells by both immunoprecipitation of metabolically labeled protein and immunohistochemical localization. We further show that externalization of the lectin is a developmentally regulated process that accompanies myoblast differentiation and that the lectin codistributes with laminin in myotube extracellular matrix. Immunohistochemical localization during intermediate stages of externalization suggests that the lectin becomes concentrated in evaginations of plasma membrane, which pinch off to form labile lectin-rich extracellular vesicles. This suggests a possible mechanism for lectin export from the cytosol to the extracellular matrix.

  3. Evidence for export of a muscle lectin from cytosol to extracellular matrix and for a novel secretory mechanism

    PubMed Central

    1990-01-01

    A soluble lactose-binding lectin with subunit Mr of 14,500 is believed to function by interacting with extracellular glycoconjugates, because it has been detected extracellularly by immunohistochemistry. This localization has been questioned, however, since the lectin lacks a secretion signal sequence, which challenges the contention that it is secreted. We have demonstrated externalization of this lectin from C2 mouse muscle cells by both immunoprecipitation of metabolically labeled protein and immunohistochemical localization. We further show that externalization of the lectin is a developmentally regulated process that accompanies myoblast differentiation and that the lectin codistributes with laminin in myotube extracellular matrix. Immunohistochemical localization during intermediate stages of externalization suggests that the lectin becomes concentrated in evaginations of plasma membrane, which pinch off to form labile lectin- rich extracellular vesicles. This suggests a possible mechanism for lectin export from the cytosol to the extracellular matrix. PMID:2335567

  4. Targeted delivery of antigen to hamster nasal lymphoid tissue with M-cell-directed lectins.

    PubMed Central

    Giannasca, P J; Boden, J A; Monath, T P

    1997-01-01

    The nasal cavity of a rodent is lined by an epithelium organized into distinct regional domains responsible for specific physiological functions. Aggregates of nasal lymphoid tissue (NALT) located at the base of the nasal cavity are believed to be sites of induction of mucosal immune responses to airborne antigens. The epithelium overlying NALT contains M cells which are specialized for the transcytosis of immunogens, as demonstrated in other mucosal tissues. We hypothesized that NALT M cells are characterized by distinct glycoconjugate receptors which influence antigen uptake and immune responses to transcytosed antigens. To identify glycoconjugates that may distinguish NALT M cells from other cells of the respiratory epithelium (RE), we performed lectin histochemistry on sections of the hamster nasal cavity with a panel of lectins. Many classes of glycoconjugates were found on epithelial cells in this region. While most lectins bound to sites on both the RE and M cells, probes capable of recognizing alpha-linked galactose were found to label the follicle-associated epithelium (FAE) almost exclusively. By morphological criteria, the FAE contains >90% M cells. To determine if apical glycoconjugates on M cells were accessible from the nasal cavity, an M-cell-selective lectin and a control lectin in parallel were administered intranasally to hamsters. The M-cell-selective lectin was found to specifically target the FAE, while the control lectin did not. Lectin bound to M cells in vivo was efficiently endocytosed, consistent with the role of M cells in antigen transport. Intranasal immunization with lectin-test antigen conjugates without adjuvant stimulated induction of specific serum immunoglobulin G, whereas antigen alone or admixed with lectin did not. The selective recognition of NALT M cells by a lectin in vivo provides a model for microbial adhesin-host cell receptor interactions on M cells and the targeted delivery of immunogens to NALT following intranasal

  5. Structure, interactions and evolutionary implications of a domain-swapped lectin dimer from Mycobacterium smegmatis.

    PubMed

    Patra, Dhabaleswar; Mishra, Padmanabh; Surolia, Avadhesha; Vijayan, Mamannamana

    2014-10-01

    Crystal structure determination of the lectin domain of MSMEG_3662 from Mycobacterium smegmatis and its complexes with mannose and methyl-α-mannose, the first effort of its kind on a mycobacterial lectin, reveals a structure very similar to β-prism II fold lectins from plant sources, but with extensive unprecedented domain swapping in dimer formation. The two subunits in a dimer often show small differences in structure, but the two domains, not always related by 2-fold symmetry, have the same structure. Each domain carries three sugar-binding sites, similar to those in plant lectins, one on each Greek key motif. The occurrence of β-prism II fold lectins in bacteria, with characteristics similar to those from plants, indicates that this family of lectins is of ancient origin and had evolved into a mature system before bacteria and plants diverged. In plants, the number of binding sites per domain varies between one and three, whereas the number is two in the recently reported lectin domains from Pseudomonas putida and Pseudomonas aeruginosa. An analysis of the sequences of the lectins and the lectin domains shows that the level of sequence similarity among the three Greek keys in each domain has a correlation with the number of binding sites in it. Furthermore, sequence conservation among the lectins from different species is the highest for that Greek key which carries a binding site in all of them. Thus, it would appear that carbohydrate binding influences the course of the evolution of the lectin. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Histological and lectin histochemical studies of the vomeronasal organ of horses.

    PubMed

    Lee, Kwang-Hyup; Park, Changnam; Kim, Jeongtae; Moon, Changjong; Ahn, Meejung; Shin, Taekyun

    2016-08-01

    The morphological characteristics and glycoconjugate composition of the vomeronasal organ (VNO) of the horse was investigated using histological, immunohistochemical, and lectin histochemical methods. The VNO is bilaterally located at the base of the nasal septum, has a tubular structure surrounded by cartilage, and consists of sensory and non-sensory epithelia. Immunohistochemical examination showed that the vomeronasal sensory epithelium (VSE) consisted of receptor cells positive for both olfactory marker protein (OMP) and protein gene product 9.5 (PGP 9.5), supporting cells, and basal cells. VNO receptor cells were positive for G protein Gαi2 (vomeronasal receptor type 1 marker), but not Gαo (vomeronasal receptor type 2 marker). Lectin histochemical studies using 21 biotinylated lectins showed that the free border of the VSE was positive for 20 lectins. The receptor and supporting cells reacted with 16 lectins while the basal cells reacted with 15 lectins, with varying intensities. In the vomeronasal non-sensory epithelium, the free border was positive for 19 lectins. The cilated cells were positive for 17 lectins and the basal cells were positive for 15 lectins. The vomeronasal glands, positioned in the lamina propria, were stained with both periodic acid Schiff (PAS) and alcian blue (pH 2.5). Eighteen lectins stained the acinar cells of the vomeronasal glands with various binding patterns. These findings suggest that horse VNO receptor cells express vomeronasal receptor type 1, and the VNO glands have mucous to seromucous characteristics. Moreover, each lectin differentially binds each cell type in both the VNO sensory and non-sensory epithelia. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Purification and characterization of a mucin specific mycelial lectin from Aspergillus gorakhpurensis: application for mitogenic and antimicrobial activity.

    PubMed

    Singh, Ram Sarup; Kaur, Hemant Preet; Singh, Jatinder

    2014-01-01

    Lectins are carbohydrate binding proteins or glycoproteins that bind reversibly to specific carbohydrates present on the apposing cells, which are responsible for their ability to agglutinate red blood cells, lymphocytes, fibroblasts, etc. Interest in lectins has been intensified due to their carbohydrate specificity as they can be valuable reagents for the investigation of cell surface sugars, purification and characterization of glycoproteins. The present study reports the purification, characterization and evaluation of mitogenic and antimicrobial potential of a mycelial lectin from Aspergillus gorakhpurensis. Affinity chromatography on mucin-sepharose column was carried out for purification of Aspergillus gorakhpurensis lectin. The lectin was characterized for physico-chemical parameters. Mitogenic potential of the lectin was evaluated against splenocytes of Swiss albino mice by MTT assay. Antimicrobial activity of the purified lectin has also been evaluated by disc diffusion assay. Single-step affinity purification resulted in 18.6-fold purification of the mycelial lectin. The molecular mass of the lectin was found to be 70 kDa and it was composed of two subunits of 34.8 kDa as determined by gel filtration chromatography, SDS-PAGE and MALDI-TOF analysis. pH optima of the lectin was found to be 6.5-9.5, while optimum temperature for lectin activity was 20-30 °C. Lectin was stable within a pH range of 7.0-10.5 and showed fair thermostability. EDTA did not affect lectin activity whereas it was found susceptible to the denaturants tested. MTT assay revealed strong mitogenic potential of A. gorakhpurensis lectin at a concentration upto 150 µg/mL. Antimicrobial activity assay showed its potent antibacterial activity against Bacillus cereus, Staphylococcous aureus and Escherichia coli and marginal antifungal activity against Saccharomyces cerevisiae. This is the first report on the mitogenic and antimicrobial potential of Aspergillus gorakhpurensis lectin. The

  8. Antinociceptive and Anti-inflammatory Activities of the Lectin from Marine Red Alga Solieria filiformis.

    PubMed

    Abreu, Ticiana Monteiro; Ribeiro, Natássia Albuquerque; Chaves, Hellíada Vasconcelos; Jorge, Roberta Jeane Bezerra; Bezerra, Mirna Marques; Monteiro, Helena Serra Azul; Vasconcelos, Ilka Maria; Mota, Érika Freitas; Benevides, Norma Maria Barros

    2016-05-01

    Lectins are proteins that bind to specific mono- or oligosaccharides. This study aimed to evaluate the antinociceptive and anti-inflammatory effects of the lectin from the red marine alga Solieria filiformis. The animals (n = 6) were pretreated with S. filiformis lectin 30 min before they were given the nociceptive or inflammatory stimulus. The antinociceptive activity was evaluated in Swiss mice using the abdominal writhing, formalin, and hot plate tests. The anti-inflammatory properties were evaluated in Wistar rats using carrageenan-induced peritonitis and paw edema induced by different phlogistic agents. The S. filiformis lectin toxicity was assayed through its application in mice (7 days). S. filiformis lectin significantly reduced the number of abdominal writhings and reduced the paw licking time in the second phase of the formalin test (p < 0.05), but it did not prolong the reaction time in the hot plate test (p > 0.05). Furthermore, S. filiformis lectin reduced neutrophil migration in a peritonitis model and reduced paw edema induced by carrageenan, dextran, and serotonin (p < 0.05). Additionally, the administration of S. filiformis lectin resulted in no signs of systemic damage. Thus, S. filiformis lectin appears to have important antinociceptive and anti-inflammatory activities and could represent a potential therapeutic agent for future studies. Georg Thieme Verlag KG Stuttgart · New York.

  9. Characterization of quail intestinal mucin as a ligand for endogenous quail lectin.

    PubMed Central

    Fang, R; Mantle, M; Ceri, H

    1993-01-01

    The S-type lectins have been shown to be components of mucosal scrapings, and in avian systems these lectins have been localized immunohistochemically to the mucosal surface and goblet cells of the intestine. The interaction of lectin specifically with purified mucin has not, however, been established. Quail intestinal mucin was purified by two subsequent isopycnic density-gradient centrifugations in CsCl and chromatography on Sepharose Cl-2B. Purified mucin, obtained from the void volume of the Sepharose column, was characterized by SDS/PAGE, amino acid and carbohydrate analyses, sensitivity to thiol reduction, and cross-reactivity with antibody preparations to rat and human intestinal mucins on Western blots. Antibody raised against purified quail mucin partially cross-reacts with purified rat, rabbit and human intestinal mucins, and specifically labels the mucosal surface and goblet cells of quail intestine by the immunoperoxidase technique. Protein eluted by lactose from an affinity matrix composed of quail intestinal mucin possessed the same molecular mass on SDS/PAGE as intestinal lectin and reacted on Western blots with a lectin-specific antibody. The data clearly demonstrate the co-localization of lectin and mucin in the quail intestine and also the ability of the lectin to specifically interact with the purified mucin, raising the question of the role of endogenous lectins in secretions. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:8352754

  10. Preparation and biological properties of a melibiose binding lectin from Bauhinia variegata seeds.

    PubMed

    Lin, Peng; Ng, Tzi Bun

    2008-11-26

    A dimeric 64-kDa melibiose-binding lectin was isolated from the seeds of Bauhinia variegata. The isolation procedure comprised affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono Q, and gel filtration on Superdex 75. The lectin was adsorbed on the first two chromatographic media. Its hemagglutinating activity was stable after 30-min exposure to temperatures up to 70 degrees C. Since lectins may demonstrate biological activities such as antiproliferative, immunomodulatory, antifungal, antiviral, and HIV-1 reverse transcriptase inhibitory activities, the isolated lectin was tested for these activities. It was found that the lectin inhibited proliferation in hepatoma HepG2 cells and breast cancer MCF7 cells with an IC(50) of 1.4 microM and 0.18 microM, respectively. HIV-1 reverse transcriptase activity was inhibited with an IC(50) of 1.02 microM. The lectin and concanavalin A (Con A) evoked maximal mitogenic response from mouse splenocytes at similar concentrations, but the maximal response to B. variegata lectin was only 1/5 of that induced by Con A in magnitude. B. variegata lectin was devoid of antifungal activity.

  11. C-type lectins do not act as functional receptors for filovirus entry into cells

    SciTech Connect

    Matsuno, Keita; Nakayama, Eri; Noyori, Osamu; Marzi, Andrea; Ebihara, Hideki; Irimura, Tatsuro; Feldmann, Heinz; Takada, Ayato

    2010-12-03

    Research highlights: {yields} Filovirus glycoprotein (GP) having a deficient receptor binding region were generated. {yields} Mutant GPs mediated virus entry less efficiently than wild-type GP. {yields} Mutant GPs bound to C-type lectins but not mediated entire steps of cellular entry. {yields} C-type lectins do not independently mediate filovirus entry into cells. {yields} Other molecule(s) are required for C-type lectin-mediated entry of filoviruses. -- Abstract: Cellular C-type lectins have been reported to facilitate filovirus infection by binding to glycans on filovirus glycoprotein (GP). However, it is not clearly known whether interaction between C-type lectins and GP mediates all the steps of virus entry (i.e., attachment, internalization, and membrane fusion). In this study, we generated vesicular stomatitis viruses pseudotyped with mutant GPs that have impaired structures of the putative receptor binding regions and thus reduced ability to infect the monkey kidney cells that are routinely used for virus propagation. We found that infectivities of viruses with the mutant GPs dropped in C-type lectin-expressing cells, parallel with those in the monkey kidney cells, whereas binding activities of these GPs to the C-type lectins were not correlated with the reduced infectivities. These results suggest that C-type lectin-mediated entry of filoviruses requires other cellular molecule(s) that may be involved in virion internalization or membrane fusion.

  12. Purification, crystallization and preliminary X-ray structure analysis of the banana lectin from Musa paradisiaca.

    PubMed

    Singh, D D; Saikrishnan, K; Kumar, Prashant; Dauter, Z; Sekar, K; Surolia, A; Vijayan, M

    2004-11-01

    The banana lectin from Musa paradisiaca, MW 29.4 kDa, has been isolated, purified and crystallized. The trigonal crystals contain one dimeric molecule in the asymmetric unit. The structure has been solved using molecular replacement to a resolution of 3 A. The structure of the subunit is similar to that of jacalin-like lectins.

  13. The roles of direct recognition by animal lectins in antiviral immunity and viral pathogenesis.

    PubMed

    Liu, Yang; Liu, Jianying; Pang, Xiaojing; Liu, Tao; Ning, Zhijie; Cheng, Gong

    2015-01-29

    Lectins are a group of proteins with carbohydrate recognition activity. Lectins are categorized into many families based on their different cellular locations as well as their specificities for a variety of carbohydrate structures due to the features of their carbohydrate recognition domain (CRD) modules. Many studies have indicated that the direct recognition of particular oligosaccharides on viral components by lectins is important for interactions between hosts and viruses. Herein, we aim to globally review the roles of this recognition by animal lectins in antiviral immune responses and viral pathogenesis. The different classes of mammalian lectins can either recognize carbohydrates to activate host immunity for viral elimination or can exploit those carbohydrates as susceptibility factors to facilitate viral entry, replication or assembly. Additionally, some arthropod C-type lectins were recently identified as key susceptibility factors that directly interact with multiple viruses and then facilitate infection. Summarization of the pleiotropic roles of direct viral recognition by animal lectins will benefit our understanding of host-virus interactions and could provide insight into the role of lectins in antiviral drug and vaccine development.

  14. Identification and characterization of C-type lectin genes from the reniform nematode

    USDA-ARS?s Scientific Manuscript database

    C-type lectins represent a large family of sugar-binding proteins which require calcium for their ligand-binding activity. C-type lectins play an important role in the innate immune response in all life forms when challenged by pathogens. Ligand binding occurs via conserved domain sequences which re...

  15. The Distribution of Lectins across the Phylum Nematoda: A Genome-Wide Search.

    PubMed

    Bauters, Lander; Naalden, Diana; Gheysen, Godelieve

    2017-01-04

    Nematodes are a very diverse phylum that has adapted to nearly every ecosystem. They have developed specialized lifestyles, dividing the phylum into free-living, animal, and plant parasitic species. Their sheer abundance in numbers and presence in nearly every ecosystem make them the most prevalent animals on earth. In this research nematode-specific profiles were designed to retrieve predicted lectin-like domains from the sequence data of nematode genomes and transcriptomes. Lectins are carbohydrate-binding proteins that play numerous roles inside and outside the cell depending on their sugar specificity and associated protein domains. The sugar-binding properties of the retrieved lectin-like proteins were predicted in silico. Although most research has focused on C-type lectin-like, galectin-like, and calreticulin-like proteins in nematodes, we show that the lectin-like repertoire in nematodes is far more diverse. We focused on C-type lectins, which are abundantly present in all investigated nematode species, but seem to be far more abundant in free-living species. Although C-type lectin-like proteins are omnipresent in nematodes, we have shown that only a small part possesses the residues that are thought to be essential for carbohydrate binding. Curiously, hevein, a typical plant lectin domain not reported in animals before, was found in some nematode species.

  16. Isolation and Characterization of Messenger RNAs for Seed Lectin and Kunitz Trypsin Inhibitor in Soybeans

    PubMed Central

    Vodkin, Lila O.

    1981-01-01

    The mRNAs for seed lectin and Kunitz trypsin inhibitor of soybean have been highly enriched by immunoadsorption of the polysomes synthesizing these proteins. Polysomes isolated from developing seed of variety Williams were incubated with monospecific rabbit antibodies produced against lectin subunits or trypsin inhibitor protein. The polysomal mixture was passed over a column containing goat anti-rabbit antibodies bound to Sepharose. Bound polysomes were eluted and the mRNA was selected by passage over oligo(dT)-cellulose. Lectin complementary DNA hybridized to an 1150-nucleotide message and trypsin inhibitor complementary DNA hybridized to a 770-nucleotide message in blotting experiments using total poly(A) RNA. Translation of soybean lectin mRNA using a rabbit reticulocyte lysate yielded a major polypeptide of 32,300 whereas the molecular weight for purified lectin subunits was 30,000. Trypsin inhibitor mRNA directed the synthesis of a 23,800-dalton polypeptide as compared to 21,500 daltons for trypsin inhibitor marker protein. Lectin specific polysomes could not be obtained from a soybean variety which lacks detectable lectin protein whereas trypsin inhibitor-specific polysomes were bound by immunoselection. These results confirmed the specificity of the immunoadsorption procedure and strongly indicated that the lectinless variety was deficient or substantially reduced in functional lectin mRNA. Images PMID:16661996

  17. Lectin-induced activation of plasma membrane NADPH oxidase in cholesterol-depleted human neutrophils.

    PubMed

    Gorudko, Irina V; Mukhortava, Ann V; Caraher, Brendan; Ren, Melody; Cherenkevich, Sergey N; Kelly, Gregory M; Timoshenko, Alexander V

    2011-12-15

    The gp91phox subunit of flavocytochrome b(558) is the catalytic core of the phagocyte plasma membrane NADPH oxidase. Its activation occurs within lipid rafts and requires translocation of four subunits to flavocytochrome b(558). gp91phox is the only glycosylated subunit of NADPH oxidase and no data exist about the structure or function of its glycans. Glycans, however, bind to lectins and this can stimulate NADPH oxidase activity. Given this information, we hypothesized that lectin-gp91phox interactions would facilitate the assembly of a functionally active NADPH oxidase in the absence of lipid rafts. To test this, we used lectins with different carbohydrate-binding specificity to examine the effects on H(2)O(2) generation by human neutrophils treated with the lipid raft disrupting agent methyl-β-cyclodextrin (MβCD). MβCD treatment removed membrane cholesterol, caused changes in cell morphology, inhibited lectin-induced cell aggregation, and delayed lectin-induced assembly of the NADPH oxidase complex. More importantly, MβCD treatment either stimulated or inhibited H(2)O(2) production in a lectin-dependent manner. Together, these results show selectivity in lectin binding to gp91phox, and provide evidence for the biochemical structures of the gp91phox glycans. Furthermore, the data also indicate that in the absence of lipid rafts, neutrophil NADPH oxidase activity can be altered by these select lectins. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Mechanism of entomotoxicity of the plant lectin from Hippeastrum hybrid (Amaryllis) in Spodoptera littoralis larvae.

    PubMed

    Caccia, Silvia; Van Damme, Els J M; De Vos, Winnok H; Smagghe, Guy

    2012-09-01

    Plant lectins have received a lot of attention because of their insecticidal properties. When orally administered in artificial diet or in transgenic plants, lectins provoke a wide range of detrimental effects, including alteration of the digestive enzyme machinery, fecundity drop, reduced feeding, changes in oviposition behavior, growth and development inhibition and mortality. Although many studies reported the entomotoxicity of lectins, only a few of them investigated the mode of action by which lectins exert toxicity. In the present paper we have studied for the first time the insecticidal potential of the plant lectin from Hippeastrum hybrid (Amaryllis) (HHA) bulbs against the larvae of the cotton leafworm (Spodoptera littoralis). Bioassays on neonate larvae showed that this mannose-specific lectin affected larval growth, causing a development retardation and larval weight decrease. Using primary cell cultures from S. littoralis midguts and confocal microscopy we have elucidated FITC-HHA binding and internalization mechanisms. We found that HHA did not exert a toxic effect on S. littoralis midgut cells, but HHA interaction with the brush border of midgut cells interfered with normal nutrient absorption in the S. littoralis midgut, thereby affecting normal larval growth in vivo. This study thus confirms the potential of mannose-specific lectins as pest control agents and sheds light on the mechanism underlying lectin entomotoxicity.

  19. Histochemical evaluation of postnatal lectin-binding sites in the mouse prostate.

    PubMed

    Sakuda, Kentaro; Muragishi, Ryoki; Yoshinaga, Kazuya

    2016-01-01

    The prostate is a male accessory genital gland that plays an essential role in reproductive function. To understand the cytological characteristics of differentiating prostatic cells, we used lectin histochemistry combined with immunohistochemistry to examine the distribution of lectin-binding sites on prostatic cells during postnatal development in the mouse. During postnatal development, Hippeastrum Hybrid Lectin (HHL) lectin reacted consistently with the luminal cells of all prostatic lobes (regions), whereas the Ricinus Communis Agglutinin I (RCA-I) and Soybean Agglutinin (SBA) lectins showed remarkable differences with age, region, and cell type. We found that the lectin-binding pattern in differentiating prostatic cells acquired adult characteristics around 3 weeks after birth. The results indicate that prostatic cell differentiation during postnatal development in mice is characterized by the presence of cell- and region-specific lectin-binding sites in the prostate, suggesting that there may also be cellular and regional differences in their function. Furthermore, some lectins (HHL, RCA-I, and SBA) could provide useful markers for research into cell differentiation and for the pathological evaluation of prostatic diseases or in the diagnosis of male infertility.

  20. Immunomodulatory and therapeutic potential of a mucin-specific mycelial lectin from Aspergillus panamensis.

    PubMed

    Singh, Ram Sarup; Kaur, Hemant Preet; Rana, Vikas; Kennedy, John F

    2017-03-01

    The present study reports the evaluation of immunomodulatory and therapeutic potential of a purified Aspergillus panamensis lectin. The immunomodulatory potential of the purified lectin was determined in swiss albino mice by studying its effect on anaphylaxis reaction, arthus reaction, respiratory burst activity, nitric oxide production and quantification of cytokine levels. The therapeutic potential of the lectin was evaluated in male wistar rat models by studying its curative effect on ulcerative colitis. The purified lectin inhibited systemic anaphylaxis and arthus reaction. It enhanced the functional ability of macrophages which was evident from increase in reduction of nitroblue tetrazolium dye and nitric oxide production. It also stimulated the production of Th-1 cytokine IFN-γ and Th-2 cytokine IL-6. Maximum immunomodulatory effect was seen at lectin concentration of 1.5mg/kg body weight. The lectin also showed curative effect against trinitrobenzene sulphonic acid induced ulcerative colitis. The results of this study adequately reflect the role of purified A. panamensis lectin in improving the immune status of mice models. They also show the effect of lectin in reducing the severity of incidence and decrease in clinical symptoms of ulcerative colitis. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Multiplex profiling of glycoproteins using a novel bead-based lectin array.

    PubMed

    Wang, Hong; Li, Hong; Zhang, Wei; Wei, Liming; Yu, Hongxiu; Yang, Pengyuan

    2014-01-01

    Lectin array is becoming important in profiling targeted glycan/glycoprotein, but weak interaction between lectin and glycan causes low sensitivity of the approach. This study aims to develop a bead-based lectin array for improving the sensitivity of glycosylation profiling. Lectins are chemically coupled to fluorescent dye coated microbeads, and glycan-lectin recognition is carried out three dimensionally. The performance of this platform was evaluated, and the LOD of lectin Ricinus communis agglutinin 120 (RCA120) was 50 pg/mL (1 pM) of asialofetuin, providing the bead-based lectin microarray with the highest sensitivity among the reported lectin microarrays. Furthermore, multiplexed assay was performed, which allowed the simultaneous detection of multiple carbohydrate epitopes in a single reaction vessel. The glycosylation patterns of hepatocellular carcinoma associated immunoglobulin G were analyzed, and increased (α-1,6) core fucosylation and (α-2,6) sialylation patterns were observed, which may provide significant clinical evidence for disease diagnosis. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Generation of ligand specificity and modes of oligomerization in β-prism I fold lectins.

    PubMed

    Chandran, Thyageshwar; Sharma, Alok; Vijayan, Mamannamana

    2013-01-01

    β-Prism I fold lectins constitute one of the five widely occurring structural classes of plant lectins. Each single domain subunit is made up of three Greek key motifs arranged in a threefold symmetric fashion. The threefold symmetry is not reflected in the sequence except in the case of the lectin from banana, a monocot, which carries two sugar-binding sites instead of the one in other lectins of known three-dimensional structure, all from dicots. This is believed to be a consequence of the different evolutionary paths followed by the lectin in monocots and dicots. The galactose-specific lectins among them have two chains produced by posttranslational proteolysis and contain three aromatic residues at the binding site. The extended binding sites of galactose- and mannose-specific lectins have been thoroughly characterized. Ligand binding at the sites involves both conformational selection and induced fit. Molecular plasticity of some of the lectins in the family has been characterized. The plasticity appears to be such as to promote variability in quaternary association which could be dimeric, tetrameric, or octameric. Structural and evolutionary reasons for the variability have been explored, and the relation of oligomerization to ligand binding and conformational selection investigated. © 2013 Elsevier Inc. All rights reserved.

  3. The Distribution of Lectins across the Phylum Nematoda: A Genome-Wide Search

    PubMed Central

    Bauters, Lander; Naalden, Diana; Gheysen, Godelieve

    2017-01-01

    Nematodes are a very diverse phylum that has adapted to nearly every ecosystem. They have developed specialized lifestyles, dividing the phylum into free-living, animal, and plant parasitic species. Their sheer abundance in numbers and presence in nearly every ecosystem make them the most prevalent animals on earth. In this research nematode-specific profiles were designed to retrieve predicted lectin-like domains from the sequence data of nematode genomes and transcriptomes. Lectins are carbohydrate-binding proteins that play numerous roles inside and outside the cell depending on their sugar specificity and associated protein domains. The sugar-binding properties of the retrieved lectin-like proteins were predicted in silico. Although most research has focused on C-type lectin-like, galectin-like, and calreticulin-like proteins in nematodes, we show that the lectin-like repertoire in nematodes is far more diverse. We focused on C-type lectins, which are abundantly present in all investigated nematode species, but seem to be far more abundant in free-living species. Although C-type lectin-like proteins are omnipresent in nematodes, we have shown that only a small part possesses the residues that are thought to be essential for carbohydrate binding. Curiously, hevein, a typical plant lectin domain not reported in animals before, was found in some nematode species. PMID:28054982

  4. Translational control of discoidin lectin expression in drsA suppressor mutants of Dictyostelium discoideum.

    PubMed Central

    Alexander, S; Leone, S; Ostermeyer, E

    1991-01-01

    Genetic analysis in Dictyostelium discoideum has identified regulatory genes which control the developmental expression of the discoidin lectin multigene family. Among these, the drsA mutation is a dominant second-site suppressor of another mutation, disB, which has the discoidinless phenotype. We now demonstrate a novel mechanism by which the drsA allele exerts its suppressive effect on the disB mutation. Interestingly, drsA does not merely bypass the disB mutation and restore the wild-type pattern of lectin expression. Rather, drsA mutant cells have high levels of discoidin lectin synthesis during growth but do not express lectins during aggregation. In contrast, wild-type cells only express lectin protein during the aggregation period of development. Phenocopies of the drsA mutation show a pattern of discoidin expression similar to that seen in the bona fide mutant. These data suggest that there may be a mechanism of negative feedback, resulting from the high levels of discoidin lectin made during growth, which inhibits further discoidin lectin expression during development. Northern (RNA) analysis of developing drsA mutant cells shows that these cells contain high levels of discoidin mRNA, although no discoidin lectin protein is being translated from these messages. Therefore, expression of the discoidin gene family can be controlled at the level of translation as well as transcription. Images PMID:2038325

  5. Assessment of weak sugar-binding ability using lectin tetramer and membrane-based glycans.

    PubMed

    Yamamoto, Kazuo

    2014-01-01

    To consider biological significance of glycosylation of proteins, it is necessary to evaluate the importance of sugar-recognition processes mediated by lectins. Though the interaction between sugars and proteins, especially animal lectins, is quite weak with K d approximately 10(-4) M, cellular and molecular recognitions mediated via sugar-protein interaction increase their avidity by 1-3 orders of magnitude by the self-association of both receptors and their ligands on cell surfaces. To assess the weak interaction between lectins and their sugar ligands, we established lectin tetramer binding to cell surface glycans using flow cytometry. This strategy is highly sensitive, and useful to determine whether or not a putative lectin domain may have sugar-binding ability.

  6. Small unilamellar vesicles as reagents: a chemically defined, quantitative assay for lectins

    SciTech Connect

    Rando, R.R.

    1981-01-01

    Samll unilamellar vesicles containing synthetic glycolipids can be prepared. These vesicles are aggregated by the appropriate lectin (Orr et al., 1979; Rando and Bangerter, 1979; Slama and Rando, 1980). It is shown here that extent of aggregation of these vesicles as measured by light scattering at 360 nm, is, under certain conditions, linear with amount of lectin added. This forms the basis of a rapid and simple quantitative assay for lectins using the modified vesicles as a defined chemical substrate. The assay is sensitive to lectin concentrations in the low ..mu..g range. The assay is applied here to studies on concanavalin A, Ricinus communis agglutinin and the ..cap alpha..-fucosyl binding lectin from Ulex europaeus (Type I).

  7. Directed evolution of lectins by an improved error-prone PCR and ribosome display method.

    PubMed

    Hu, Dan; Tateno, Hiroaki; Hirabayashi, Jun

    2014-01-01

    Lectins are useful reagents for the structural characterization of glycans. However, currently available lectins have an apparent drawback in their "repertoire," lacking some critical probes, such as those for sulfated glycans. Thus, engineering lectins with novel specificity would be of great practical value. Here, we describe a directed evolution strategy to tailor novel lectins for novel specificity or biological functions. Our strategy uses a reinforced ribosome display-based selection combined with error-prone PCR to isolate mutants with target specificity and an evanescent-field fluorescence-assisted glycoconjugate microarray to rapidly evaluate the specificity of selected mutants. A successful case of screening a lectin, which has acquired an ability to recognize 6-sulfo-galactose-terminated glycans, is described.

  8. Correlation of Interleukin-6 levels and lectins during Schistosoma haematobium infection.

    PubMed

    Antony, Justin S; Ojurongbe, Olusola; Meyer, Christian G; Thangaraj, Kumarasamy; Mishra, Anshuman; Kremsner, Peter G; Velavan, Thirumalaisamy P

    2015-12-01

    Urogenital schistosomiasis caused by Schistosoma haematobium induces a Th2 immune response, including expression of Interleukin-6. IL-6 confers protection from experimental Schistosoma-induced pulmonary hypertension and modulates production of mannose-binding lectin (MBL) and other lectins. We studied IL-6 levels in schistosomiasis and its effect on lectins production. Elevated IL-6 levels occurred in cases, compared to controls. IL-6 correlated with the lectins MBL, ficolin-2 and Collectin Kidney-1 (CL-K1) in cases, but correlated inversely in controls. The study shows that IL-6 levels are elevated in individuals infected with urogenital schistosomiasis. IL-6 was also found to be correlated with the production of lectins in S. haematobium infection. A similar correlation between IL-6 and MBL was observed during visceral leishmaniasis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Developmental changes in the distribution of cecal lectin-binding sites of Balb-c mice.

    PubMed

    Doehrn, S; Breipohl, W; Lierse, W; Romaniuk, K; Young, W

    1992-01-01

    The existence of lectin-binding sites was investigated in the cecum of Balb-c mice at seven developmental stages ranging from 18 days post conception (p.c.) to 8 weeks after birth. Nine horseradish-peroxidase-conjugated lectins (concanavalin A, Triticum vulgaris, Dolichus biflorus, Helix pomatia, Arachis hypogaea, Glycine maximus, Lotus tetragonolobus, Ulex europaeus, Limulus polyphemus) were applied to 5- to 7-microns thin paraffin sections of Bouin-fixed tissue. After DAB staining the sections were evaluated by light microscopy. It was shown that each lectin exhibits a unique developmental pattern. The adult binding patterns were established at the age of 3-4 weeks with only minor changes occurring thereafter. Considerable differences in binding patterns occurred not only between lectins of different groups but also between lectins with the same nominal monosaccharide specificity.

  10. Deterrent activity of plant lectins on cowpea weevil Callosobruchus maculatus (F.) oviposition.

    PubMed

    Sadeghi, Amin; Van Damme, Els J M; Peumans, Willy J; Smagghe, Guy

    2006-09-01

    A set of 14 plant lectins was screened in a binary choice bioassay for inhibitory activity on cowpea weevil Callosobruchus maculatus (F.) oviposition. Coating of chickpea seeds (Cicer arietinum L.) with a 0.05% (w/v) solution of plant lectins caused a significant reduction in egg laying. Control experiments with heat inactivated lectin and BSA indicated that the observed deterrent effects are specific and require carbohydrate-binding activity. However, no clear correlation could be established between deterrent activity and sugar-binding specificity/molecular structure of the lectins. Increasing the insect density reduced the inhibitory effect of the lectins confirming that female insects are capable of adjusting their oviposition rates as a function of host availability.

  11. High-resolution crystal structures of Colocasia esculenta tarin lectin.

    PubMed

    Pereira, Patricia R; Meagher, Jennifer L; Winter, Harry C; Goldstein, Irwin J; Paschoalin, Vânia M F; Silva, Joab T; Stuckey, Jeanne A

    2017-01-01

    Tarin, the Colocasia esculenta lectin from the superfamily of α-d-mannose-specific plant bulb lectins, is a tetramer of 47 kDa composed of two heterodimers. Each heterodimer possesses homologous monomers of ~11.9 (A chain) and ~12.7 (B chain) kDa. The structures of apo and carbohydrate-bound tarin were solved to 1.7 Å and 1.91 Å, respectively. Each tarin monomer forms a canonical β-prism II fold, common to all members of Galanthus nivalis agglutinin (GNA) family, which is partially stabilized by a disulfide bond and a conserved hydrophobic core. The heterodimer is formed through domain swapping involving the C-terminal β-strand and the β-sheet on face I of the prism. The tetramer is assembled through the dimerization of the B chains from heterodimers involving face II of each prism. The 1.91 Å crystal structure of tarin bound to Manα(1,3)Manα(1,6)Man reveals an expanded carbohydrate-binding sequence (QxDxNxVxYx4/6WX) on face III of the β-prism. Both monomers possess a similar fold, except for the length of the loop, which begins after the conserved tyrosine and creates the binding pocket for the α(1,6)-terminal mannose. This loop differs in size and amino-acid composition from 10 other β-prism II domain proteins, and may confer carbohydrate-binding specificity among members of the GNA-related lectin family.

  12. Chemical characteristics of dimer interfaces in the legume lectin family.

    PubMed

    Elgavish, S; Shaanan, B

    2001-04-01

    The Erythrina corallodendron lectin (EcorL) crystallizes in monoclinic and hexagonal crystal forms. Comparison of the newly determined hexagonal form (PDB code 1fyu) with the monoclinic form shows that the dimeric structure of EcorL reflects the inherent biological structure of the protein and is not an artifact of the crystal packing. To further understand the factors determining the dimerization modes of legume lectins, EcorL, concanavalin A (ConA), and Griffonia simplicifolia (GS4) were taken as representatives of the three unique dimers found in the family. Six virtual homodimers were generated. The hydropathy, amino acid composition, and solvation energy were calculated for all nine homodimers. Each of the three native dimers has a distinct chemical composition. EcorL has a dominant hydrophobic component, and ConA has a strong polar component, but in GS4 the three components contribute equally to the interface. This distribution pattern at the interface is unique to the native dimers and distinct from the partition observed in the virtual dimers. Amino acid composition of other members of the family that dimerize like EcorL or ConA maintain the same pattern of amino acids distribution observed in EcorL and ConA. However, lectins that dimerize like GS4 do not show a particularly distinct distribution. In all cases, the calculated solvation energy of the native dimer was lower than that of the virtual dimers, suggesting that the observed mode of dimerization is the most stable organization for the given sequence and tertiary structure. The dimerization type cannot be predicted by sequence analysis.

  13. Adaptive Evolution of a Novel Drosophila Lectin Induced by Parasitic Wasp Attack

    PubMed Central

    Keebaugh, Erin S.; Schlenke, Todd A.

    2012-01-01

    Drosophila melanogaster has long been used as a model for the molecular genetics of innate immunity. Such work has uncovered several immune receptors that recognize bacterial and fungal pathogens by binding unique components of their cell walls and membranes. Drosophila also act as hosts to metazoan pathogens such as parasitic wasps, which can infect a majority of individuals in natural populations, but many aspects of their immune responses against these more closely related pathogens are poorly understood. Here, we present data describing the transcriptional induction and molecular evolution of a candidate Drosophila anti-wasp immunity gene, lectin-24A. Lectin-24A has a secretion signal sequence and its lectin domain suggests a function in sugar group binding. Transcript levels of lectin-24A were induced significantly stronger and faster following wasp attack than following wounding or bacterial infection, demonstrating lectin-24A is not a general stress response or defense response gene but is instead part of a specific response against wasps. The major site of lectin-24A transcript production is the fat body, the main humoral immune tissue of flies. Interestingly, lectin-24A is a new gene of the D. melanogaster/Drosophila simulans clade, displaying very little homology to any other Drosophila lectins. Population genetic analyses of lectin-24A DNA sequence data from African and North American populations of D. melanogaster and D. simulans revealed gene length polymorphisms segregating at high frequencies as well as strong evidence of repeated and recent selective sweeps. Thus, lectin-24A is a rapidly evolving new gene that has seemingly developed functional importance for fly resistance against infection by parasitic wasps. PMID:21873297

  14. Isolation and characterization of a novel lectin with mitogenic activity from Pleurotus ferulae.

    PubMed

    Xu, Cheng-Jian; Wang, Yue-Xiang; Niu, Bo-Nan; Liu, Bing; Li, Ying-Biao; Wang, Xue-Ming; Lu, Shi-Ling

    2014-07-01

    Lectins are the tools for the determination of sugar chain structure. Recently, lectin arrays have become a popular new technology; therefore, lectins with specific sugar-binding properties are required. The objective of the study was to isolate a novel lectin from Pleurotus ferulae mushrooms and characterize its various biological activities. A novel lectin was extracted with deionized water, precipitated from the aqueous extract using 75% saturated (NH4)2SO4, and subjected on DEAE-cellulose followed by affinity chromatography on sepharose-6B. The activity was tested using hemagglutination assays, and carbohydrate-binding specificity was determined by glycan microarray analysis. Its effects on the mitogenic activity of mouse splenocytes were determined by MTT assay. The novel lectin was adsorbed on ion-exchange chromatography DEAE-cellulose and shown as a band with the molecular mass of 17.5 kDa on a SDS-PAGE and as a single 35.0-kDa peak in gel filtration on Superdex G-75. The hemagglutinating activity of the lectin was inhibited by D-glucose, lactose, D-galactose, and galactosamine. The lectin was stable on 60°C. The hemagglutinating activity of lectin was reduced by 50% at 70°C. At 80°C, it was further reduced to 6.25% of its original activity. The hemagglutinating activity was the highest at pH 6-9. Moreover, its hemagglutinating activity was inhibited by Mg2+ and Ca2+ ions. The lectin isolated from P. ferulae in the current study possessed highly potent hemagglutinating and proliferative activities toward mouse splenocytes.

  15. Lectin conjugates as potent, nonabsorbable CFTR inhibitors for reducing intestinal fluid secretion in cholera.

    PubMed

    Sonawane, N D; Zhao, Dan; Zegarra-Moran, Olga; Galietta, Luis J V; Verkman, A S

    2007-04-01

    Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel are predicted to prevent intestinal fluid secretion in cholera. We previously discovered low- affinity glycine hydrazide (GlyH) CFTR inhibitors that block CFTR at its external pore. The goal of this study was to develop potent CFTR inhibitors that are minimally absorbed and washed out of the intestinal lumen for application as antisecretory agents in cholera. GlyH analogs (malonic hydrazides, MalH) were chemically conjugated to various lectins ("MalH-lectin") and purified. CFTR inhibition potency was measured by short-circuit current analysis, mechanism of action by patch-clamp, and antidiarrheal efficacy in closed-loop and suckling mouse models. By lectin conjugation, we improved CFTR inhibitory potency by approximately 100-fold (to 50 nmol/L) and retarded washout. High-affinity CFTR inhibition was abolished by MalH-lectin heat denaturation, protease digestion, or competition by mannose or unconjugated lectin. Patch-clamp analysis indicated CFTR inhibition by an external pore occlusion mechanism. Fluorescently labeled MalH-lectin remained membrane bound for >6 hours after washout, whereas washout occurred in a few minutes without the lectin. MalH-ConA and MalH-wheat (IC50 50-100 pmol) blocked cholera toxin-induced intestinal fluid secretion in closed intestinal loops in mice and greatly reduced mortality in a suckling mouse model of cholera. The high potency of MalH-lectin conjugates results from "anchoring" the CFTR-blocking MalH to cell surface carbohydrates by the lectin. The high-affinity, slow washout, and external site of action of the MalH-lectin conjugates support their further development as antisecretory drugs for enterotoxin-mediated secretory diarrheas.

  16. Microscopic and biochemical characterization of lectin binding sites in the cephalopod retina.

    PubMed

    Taba, A; Quezada, B H; Robles, L J

    1989-05-22

    Using light and electron microscope cytochemistry and lectin blotting techniques, we have shown that the lectins concanavalin A (Con A), Ricinus communis agglutinin (RCA), and peanut agglutinin (PNA) bind to specific glycoconjugants in the adult cephalopod retina. For light microscope lectin cytochemistry, aldehyde-fixed, frozen, or Araldite-embedded, etched sections of cephalopod retinas were incubated with FITC- or TRITC-conjugated lectins and examined by using epifluorescence microscopy. Con A labeled structures in the entire retina including the inner limiting membrane (ILM), rhabdomeric membranes, interphotoreceptor matrix (IPM), and structures in the photoreceptor inner segments. RCA labeling was similar to that of Con A except that there was a decrease in the staining of the rhabdom tips near the ILM. PNA labeled only the interphotoreceptor matrix between apposing rhabdomeres. The intensity of staining of the IPM by PNA also decreased or was absent toward the rhabdom tips. None of the lectins labeled the myeloid bodies located in the photoreceptor inner segments. Electron microscope (EM) lectin cytochemistry was performed on aldehyde-fixed, LR White-embedded tissue or on Araldite-embedded, periodate-etched sections by using gold-conjugated lectins. EM results confirmed the observations made by light microscopy. Lectin blots with a retinal extract or light-sensitive membrane fraction revealed a variety of protein bands labeled by all three lectins. Con A and RCA labeled opsin and its aggregates whereas PNA did not. None of the lectins labeled retinochrome. The labeling of the cephalopod IPM by PNA suggests a structural similarity between the IPM of vertebrates and invertebrates. In other studies, we have demonstrated the presence of a retinoid binding protein in the IPM of cephalopods.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Adaptive evolution of a novel Drosophila lectin induced by parasitic wasp attack.

    PubMed

    Keebaugh, Erin S; Schlenke, Todd A

    2012-02-01

    Drosophila melanogaster has long been used as a model for the molecular genetics of innate immunity. Such work has uncovered several immune receptors that recognize bacterial and fungal pathogens by binding unique components of their cell walls and membranes. Drosophila also act as hosts to metazoan pathogens such as parasitic wasps, which can infect a majority of individuals in natural populations, but many aspects of their immune responses against these more closely related pathogens are poorly understood. Here, we present data describing the transcriptional induction and molecular evolution of a candidate Drosophila anti-wasp immunity gene, lectin-24A. Lectin-24A has a secretion signal sequence and its lectin domain suggests a function in sugar group binding. Transcript levels of lectin-24A were induced significantly stronger and faster following wasp attack than following wounding or bacterial infection, demonstrating lectin-24A is not a general stress response or defense response gene but is instead part of a specific response against wasps. The major site of lectin-24A transcript production is the fat body, the main humoral immune tissue of flies. Interestingly, lectin-24A is a new gene of the D. melanogaster/Drosophila simulans clade, displaying very little homology to any other Drosophila lectins. Population genetic analyses of lectin-24A DNA sequence data from African and North American populations of D. melanogaster and D. simulans revealed gene length polymorphisms segregating at high frequencies as well as strong evidence of repeated and recent selective sweeps. Thus, lectin-24A is a rapidly evolving new gene that has seemingly developed functional importance for fly resistance against infection by parasitic wasps.

  18. Dietary garlic (Allium sativum) lectins, ASA I and ASA II, are highly stable and immunogenic.

    PubMed

    Clement, Fatima; Venkatesh, Yeldur P

    2010-10-01

    The immunomodulatory proteins present in garlic have recently been shown to be identical to the garlic lectins ASA I and ASA II [Clement F, Pramod SN, Venkatesh YP. Int. Immunopharmacol. 2010; 10: 316-324]. In this study, the stability of garlic lectins as a function of pH, temperature and denaturants has been examined in relation to biological activity (hemagglutination and hagocytosis). Stability of garlic lectins in simulated gastric fluid (SGF) was assessed by their hemagglutination activity, immunoreactivity, and intactness by SDS-PAGE. Garlic lectins were moderately stable in SGF for up to 30 min; while they retained hemagglutination activities, immunoreactivity with the respective rabbit antiserum decreased immediately (0.5 min) to 10-30%. ASA I retained ~80% hemagglutination activity in the pH range 2-12; however, ASA II retained only 40% in the pH ranges 2-4 and 10-12. Garlic lectins exposed to 60 °C (30 min) and pepsin (1 and 2 min) retained hemagglutination and phagocytic activities. Urea (4M) and Gdn.HCl (2M) did not affect hemagglutination. The immunogenicity of garlic lectins upon oral feeding in BALB/c mice was examined. A lectin-specific serum IgG response was seen in mice comparable to the oral immunogen, phytohemagglutinin. The recovered lectin in feces of mice administered with garlic lectins showed antigenicity identical to that of the administered proteins. The stabilities of the garlic lectins, their ability to withstand the gastrointestinal passage, and their recognition by the immune system upon oral feeding reinforce the reported presence of natural antibodies to garlic proteins in normal human sera.

  19. Lectin-based Isolation and Culture of Mouse Embryonic Motoneurons

    PubMed Central

    Conrad, Rebecca; Jablonka, Sibylle; Sczepan, Teresa; Sendtner, Michael; Wiese, Stefan; Klausmeyer, Alice

    2011-01-01

    Spinal motoneurons develop towards postmitotic stages through early embryonic nervous system development and subsequently grow out dendrites and axons. Neuroepithelial cells of the neural tube that express Nkx6.1 are the unique precursor cells for spinal motoneurons1. Though postmitotic motoneurons move towards their final position and organize themselves into columns along the spinal tract2,3. More than 90% of all these differentiated and positioned motoneurons express the transcription factors Islet 1/2. They innervate the muscles of the limbs as well as those of the body and the inner organs. Among others, motoneurons typically express the high affinity receptors for brain derived neurotrophic factor (BDNF) and Neurotrophin-3 (NT-3), the tropomyosin-related kinase B and C (TrkB, TrkC). They do not express the tropomyosin-related kinase A (TrkA)4. Beside the two high affinity receptors, motoneurons do express the low affinity neurotrophin receptor p75NTR. The p75NTR can bind all neurotrophins with similar but lower affinity to all neurotrophins than the high affinity receptors would bind the mature neurotrophins. Within the embryonic spinal cord, the p75NTR is exclusively expressed by the spinal motoneurons5. This has been used to develop motoneuron isolation techniques to purify the cells from the vast majority of surrounding cells6. Isolating motoneurons with the help of specific antibodies (panning) against the extracellular domains of p75NTR has turned out to be an expensive method as the amount of antibody used for a single experiment is high due to the size of the plate used for panning. A much more economical alternative is the use of lectin. Lectin has been shown to specifically bind to p75NTR as well7. The following method describes an alternative technique using wheat germ agglutinin for a preplating procedure instead of the p75NTR antibody. The lectin is an extremely inexpensive alternative to the p75NTR antibody and the purification grades using

  20. Re-examining the proposed lectin properties of IL-2.

    PubMed

    Papalia, Giuseppe A; Rini, James M

    2008-03-01

    Early work examining the interactions of IL-2 and the urinary glycoprotein uromodulin led to the suggestion that IL-2 was a lectin with specificity for high-mannose and mannan ligands. Subsequent studies have attributed various roles to these properties, some critical to the cell proliferative activity of IL-2. In an attempt to verify the reported interaction between IL-2 and mannose containing carbohydrate ligands we studied two biologically active forms of IL-2 using various techniques including affinity chromatography, equilibrium dialysis, and NMR methods. Despite previous reports we have not been able to demonstrate that IL-2 possesses the ability to bind carbohydrate.

  1. Lectin sensitized anisotropic silver nanoparticles for detection of some bacteria.

    PubMed

    Gasparyan, Vardan K; Bazukyan, Inga L

    2013-03-05

    A method of bacteria detection by sensitized anisotropic silver nanoparticles is presented. Anisotropic silver nanoparticles with two bands of surface plasmon resonance (SPR) are prepared and sensitized with potato lectin. These nanoparticles are able to detect three bacterial species: Escherichia coli, Bacillus subtilis and Staphylococcus aureus. The interaction of these bacteria with such nanoparticles induces drastic changes in optical spectra of nanoparticles that are correlated with bacteria titer. The maximal sensitivity is observed for S. aureus (up to 1.5×10(4) mL(-1)).

  2. Probing lectin and sperm with carbohydrate-modified quantum dots.

    PubMed

    Robinson, Anandakathir; Fang, Jim-Min; Chou, Pi-Tai; Liao, Kuang-Wen; Chu, Rea-Min; Lee, Shyh-Jye

    2005-10-01

    We report the encapsulation of quantum dots with biologically important beta-N-acetylglucosamine (GlcNAc) in different ratios, together with studies of their specific/sensitive multivalent interactions with lectins and sperm by fluorimetry, transmission electron microscopy, dynamic light scattering microscopy, confocal imaging techniques, and flow cytometry. These GlcNAc-encapsulated quantum dots (QDGLNs) specifically bind to wheat germ agglutinin, and cause fluorescence quenching and aggregation. Further studies of QDGLNs and the mannose-encapsulated QDs (QDMANs) with sperm revealed site-specific interactions, in which QDGLNs bind to the head of the sperm, while QDMANs spread over the whole sperm body.

  3. Analysis of EST and lectin expressions in hemocytes of Manila clams (Ruditapes philippinarum) (Bivalvia: Mollusca) infected with Perkinsus olseni.

    PubMed

    Kang, Yoon-Suk; Kim, Young-Mee; Park, Kyung-Il; Kim Cho, Somi; Choi, Kwang-Sik; Cho, Moonjae

    2006-01-01

    The hemocytes of invertebrates play key roles in both cellular and humoral immune reactions by phagocytosis or delivering immune factors such as lectin and anti-microbial peptides. Bacterial infection causes changes in components such as lectins, anti-bacterial peptides, and lysosomal enzymes of plasma or hemolymph in molluscs. Previously, we found that infection with the protozoan parasite, Perkinsus, increases lectin synthesis in hemocytes. In order to investigate the patterns of genes expressed in Manila clams (Ruditapes philippinarum) infected with the protozoan parasite Perkinsus olseni, we constructed a cDNA library and sequenced 1850 clones (expressed sequence tags). A total of 79 ESTs, were related to 29 functional immune genes such as C-type lectin, lysozyme, and cystatin B, in Manila clams. Lectins were the largest group of immune-function ESTs found in our Manila clams library. Among 7 lectin clones, two full length cDNAs of lectins were cloned. MCL-3, which is a simple C-type lectin composed of 151 amino acids, has a relatively short signal sequence of 17aa and single carbohydrate-recognition domain (CRD) of approximately 130 residues. It is highly homologous to eel C-type lectin. The sequence of mc-sialic acid-binding lectin consists of 168 amino acid residues with molecular weight of 19.2 and shows high homology to sialic acid-binding lectin from the snail, Cepaea hortensis. The expression of 7 different lectins in hemocytes was analyzed by RT-PCR using gene-specific primers. Hemocytes from Perkinsus-infected clam expressed different sets of lectins than with Vibrio infection. These results demonstrate that several lectins are involved in Manila clam innate immunity and different challenges induce expression of different lectins.

  4. Extraction and purification of a lectin from red kidney bean and preliminary immune function studies of the lectin and four Chinese herbal polysaccharides.

    PubMed

    Hou, Yufang; Hou, Yubao; Yanyan, Liu; Qin, Guang; Li, Jichang

    2010-01-01

    Reversed micelles were used to extract lectin from red kidney beans and factors affecting reverse micellar systems (pH value, ionic strength and extraction time) were studied. The optimal conditions were extraction at pH 4-6, back extraction at pH 9-11, ion strength at 0.15 M NaCl, extraction for 4-6 minutes and back extraction for 8 minutes. The reverse micellar system was compared with traditional extraction methods and demonstrated to be a time-saving method for the extraction of red kidney bean lectin. Mitogenic activity of the lectin was reasonably good compared with commercial phytohemagglutinin (extracted from Phaseolus vulgaris) Mitogenic properties of the lectin were enhanced when four Chinese herbal polysaccharides were applied concurrently, among which 50 μg/mL Astragalus mongholicus polysaccharides (APS) with 12.5 μg/mL red kidney bean lectin yielded the highest mitogenic activity and 100 mg/kg/bw APS with 12.5 mg/kg/bw red kidney bean lectin elevated mouse nonspecific immunity.

  5. Extraction and Purification of a Lectin from Red Kidney Bean and Preliminary Immune Function Studies of the Lectin and Four Chinese Herbal Polysaccharides

    PubMed Central

    Hou, Yufang; Hou, Yubao; Yanyan, Liu; Qin, Guang; Li, Jichang

    2010-01-01

    Reversed micelles were used to extract lectin from red kidney beans and factors affecting reverse micellar systems (pH value, ionic strength and extraction time) were studied. The optimal conditions were extraction at pH 4–6, back extraction at pH 9–11, ion strength at 0.15 M NaCl, extraction for 4–6 minutes and back extraction for 8 minutes. The reverse micellar system was compared with traditional extraction methods and demonstrated to be a time-saving method for the extraction of red kidney bean lectin. Mitogenic activity of the lectin was reasonably good compared with commercial phytohemagglutinin (extracted from Phaseolus vulgaris) Mitogenic properties of the lectin were enhanced when four Chinese herbal polysaccharides were applied concurrently, among which 50 μg/mL Astragalus mongholicus polysaccharides (APS) with 12.5 μg/mL red kidney bean lectin yielded the highest mitogenic activity and 100 mg/kg/bw APS with 12.5 mg/kg/bw red kidney bean lectin elevated mouse nonspecific immunity. PMID:20976304

  6. High-resolution structure of a new Tn antigen-binding lectin from Vatairea macrocarpa and a comparative analysis of Tn-binding legume lectins.

    PubMed

    Sousa, Bruno Lopes; Silva Filho, José Caetano; Kumar, Prashant; Pereira, Ronniery Ilário; Łyskowski, Andrzej; Rocha, Bruno Anderson Matias; Delatorre, Plínio; Bezerra, Gustavo Arruda; Nagano, Celso Shiniti; Gruber, Karl; Cavada, Benildo Sousa

    2015-02-01

    Plant lectins have been studied as histological markers and promising antineoplastic molecules for a long time, and structural characterization of different lectins bound to specific cancer epitopes has been carried out successfully. The crystal structures of Vatairea macrocarpa (VML) seed lectin in complex with GalNAc-α-O-Ser (Tn antigen) and GalNAc have been determined at the resolution of 1.4Å and 1.7Å, respectively. Molecular docking analysis of this new structure and other Tn-binding legume lectins to O-mucin fragments differently decorated with this antigen provides a comparative binding profile among these proteins, stressing that subtle alterations that may not influence monosaccharide binding can, nonetheless, directly impact the ability of these lectins to recognize naturally occurring antigens. In addition to the specific biological effects of VML, the structural and binding similarities between it and other lectins commonly used as histological markers (e.g., VVLB4 and SBA) strongly suggest VML as a candidate tool for cancer research. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Effects of lectins with various carbohydrate binding specificities on lipid metabolism in isolated rat and hamster adipocytes.

    PubMed

    Ng, T B; Li, W W; Yeung, H W

    1989-01-01

    1. Mannose-binding and N-acetylglucosamine binding lectins exhibited potent antilipolytic and lipogenic activities. 2. Fucose-binding lectins had minimal lipogenic activity but possessed antilipolytic activity. 3. Most galactose-binding and N-acetylgalactosamine-binding lectins were devoid of significant antilipolytic and lipogenic activities. Notable exceptions were lectins from Momordica charantia, Wisteria floribunda, Vicia villosa, Codium fragile and the Siberian pine tree. lipogenic activity but lacked antilipolytic activity. 5. The galactose-binding horse gram and osage orange lectins exerted antilipolytic activity in hamster but not in rat adipocytes.

  8. Two novel lectins from Parkia biglandulosa and Parkia roxburghii: isolation, physicochemical characterization, mitogenicity and anti-proliferative activity.

    PubMed

    Kaur, Navjot; Singh, Jatinder; Kamboj, Sukhdev Singh; Agrewala, Javed N; Kaur, Manpreet

    2005-08-01

    Two mannose/glucose specific seed lectins were isolated from Parkia biglandulosa and Parkia roxburghii and were characterized w.r.t various physicochemical properties. Unlike other Parkia lectins a comparison of native and subunit molecular mass showed that both Parkia lectins were heterotetramers. Parkia biglandulosa lectin was found to be T-cell mitogen as revealed by IL-2 bioassay. These lectins showed anti-proliferative effect on two murine macrophage cancer cell lines i.e. P 388DI (50%) and J774 (70%). In addition Parkia roxburghii also inhibited proliferation of HB98 (65.47%), a B-cell hybridoma cell line.

  9. Gal/GalNAc specific multiple lectins in marine bivalve Anadara granosa.

    PubMed

    Adhya, Mausumi; Singha, Biswajit

    2016-03-01

    Complete lectin mapping of molluscs with their diversified recognition pattern and possible role in lectin-carbohydrate interaction based immune response triggering need much attention. In this communication, Gal/GalNAc specific three lectins AGL-IA (Anadara granosa lectin-IA), AGL-IB (A. granosa lectin-IB) and AGL-IV (A. granosa lectin-IV) and a lectin having hemolytic activity AGL-III (A. granosa lectin-III) were purified from the plasma of A. granosa bivalve by a combination of gel filtration and affinity chromatography. AGL-IA and IB were oligomeric lectins whereas, AGL-III and IV were monomeric. The molecular weight of AGL-IA, IB, III and IV were 375, 260, 45 and 33 kDa respectively. AGL-IA and IV agglutinated both rabbit and pronase treated human erythrocytes, whereas AGL-IB agglutinated only rabbit erythrocytes. AGL-III was found to agglutinate rabbit erythrocytes, however, it caused hemolysis of pronase treated human erythrocytes. The activity of all four lectins was calcium dependent and maximum at a pH range 7-8. Apart from Gal/GalNAc specific, the four lectins showed substantial differences in their carbohydrate recognition pattern. Moreover, there was a difference in the carbohydrate specificity between AGL-III and other three lectins (AGL-IA, AGL-IB and AGL-IV) towards polyvalent glycotope. On the one hand, 'cluster glycoside effect' i.e., an enhancement of the activity of a multivalent ligand, was observed for carbohydrate specificities of AGL-IA, AGL-IB, AGL-IV. On the other hand, the effect of multivalent ligands on the carbohydrate specificity of AGL-III was opposite of cluster glycoside effect. The affinity of AGL-IA, AGL-IB and AGL-IV for ligands can be ranked as follows: glycoproteins > polysaccharide > oligosaccharides and monosaccharides. However, Gal related monosaccharides were the best inhibitors of AGL-III and the inhibitory activity decreased gradually in the following order: monosaccharide > disaccharide > polysaccharide. Thus, the

  10. Purification and Characterization of Two Major Lectins from Araucaria brasiliensis syn. Araucaria angustifolia Seeds (Pinhão) 1

    PubMed Central

    Datta, Pradip K.; Figueroa, Maria O. D. C. R.; Lajolo, Franco M.

    1991-01-01

    Two major lectins (lectin I and lectin II) were purified to homogeneity from the seeds of Araucaria brasiliensis (Gymnospermae). The purity of the lectins was confirmed by polyacrylamide gel electrophoresis, isoelectric focusing, and high performance liquid chromatography. They are glycoproteins in nature containing 6.3 and 2.9%, respectively, of neutral sugar and have absorption coefficients of 3.8 and 4.7, respectively, at 280 nanometers. The molecular weights of both lectins obtained by gel filtration on Sephacryl S-400 were equal: 200,000. After dissociation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, molecular weights were 20,000 and 34,000, respectively, for lectin I and lectin II, suggesting they are decameric and hexameric in nature. The amino acid composition of both lectins showed little difference, but both had high amounts of acidic amino acids and lacked methionine in their molecule. The carbohydrate binding specificity of lectins was directed towards mannose, glucose, and their oligomers. High inhibitory activity was also found with thyroglobulin. The erythroagglutinating activity of the lectins was enhanced in the presence of high-molecular-weight substances both at 37 and 4°C. Divalent cations do not appear to be essential for activity. They maintained their agglutinating activity over a broad but different range of pH: 5.5 to 7.5 and 6.5 to 7.5, respectively. Both lectins agglutinated erythrocytes of human ABO blood types equally well. ImagesFigure 2Figure 3 PMID:16668523

  11. The degradation of lectins, phaseolin and trypsin inhibitors during germination of white kidney beans, Phaseolus vulgaris L.

    PubMed

    Savelkoul, F H; Tamminga, S; Leenaars, P P; Schering, J; Ter Maat, D W

    1994-04-01

    White kidney beans (Phaseolus vulgaris), cv Processor, contain a relatively high content of phaseolin (storage protein), lectins and a special group of glycoproteins as well as a considerable amount of protein-type trypsin inhibitors. Protein digestion of raw 'Processor' beans in monogastrics, for example pigs, is disturbed by poorly digested, phaseolin lectins, which can bind to carbohydrates in brush border membranes of the small intestinal epithelium, and trypsin inhibitors. The effect of the germination of white kidney beans on lectins, phaseolin and trypsin inhibitors was studied in order to achieve a degradation of lectins, phaseolin and trypsin inhibitors and an increase of in vitro enzymatic hydrolysis of the protein of bean flour. Therefore, whole bean extracts were examined throughout a germination period of up to seven days for their lectin and phaseolin pattern, lectin content, binding capacities of functional lectins towards brush border membranes and trypsin inhibitor content. In addition the in vitro enzymatic hydrolysis by pepsin and pancreatin of the protein from flours of (un)germinated white kidney beans was studied. SDS-PAGE demonstrated a degradation of E-lectins and a disappearance of L-lectins and phaseolin during germination. Results indicated a decrease of the lectin content by 85%, a loss of binding capacities of functional lectins towards brush border membranes by 91%, and a decrease of trypsin inhibitors by 76%, in bean flour after germination for seven days. A maximum in in vitro enzymatic hydrolysis of protein from bean flour was already established after germination for half a day.

  12. Lectin functionalized quantum dots for recognition of mammary tumors

    NASA Astrophysics Data System (ADS)

    Santos, Beate S.; de Farias, Patricia M. A.; de Menezes, Frederico D.; de C. Ferreira, Ricardo; Júnior, Severino A.; Figueiredo, Regina C. B. Q.; Beltrão, Eduardo I. C.

    2006-02-01

    In this study we use CdS/Cd(OH) II quantum dots functionalized with concanavalin-A (Con-A) lectin, specific to glucose/mannose residues, to investigate cell alterations regarding carbohydrate profile in human mammary tissues diagnosed as fibroadenoma (benign tumor). These particles were functionalized with glutaraldehyde and Con-A and incubated with tissue sections of normal and to Fibroadenoma, a benign type of mammary tumor. The tissue sections were deparafinized, hydrated in graded alcohol and treated with a solution of Evans Blue in order to avoid autofluorescence. The fluorescence intensity of QD-Con-A stained tissues showed different patterns, which reflect the carbohydrate expression of glucose/mannose in fibroadenoma when compared to the detection of the normal carbohydrate expression. The pattern of unspecific labeling of the tissues with glutaraldehyde functionalized CdS/Cd(OH) II quantum dots is compared to the targeting driven by the Con-A lectin. The preliminary findings reported here support the use of CdS/Cd(OH) II quantum dots as specific probes of cellular alterations and their use in diagnostics.

  13. Detection of colorectal dysplasia using fluorescently labelled lectins

    PubMed Central

    Kuo, Joe Chin-Hun; Ibrahim, Ashraf E. K.; Dawson, Sarah; Parashar, Deepak; Howat, William J.; Guttula, Kiran; Miller, Richard; Fearnhead, Nicola S.; Winton, Douglas J.; Neves, André A.; Brindle, Kevin M.

    2016-01-01

    Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions. We tested here the ability of fluorescently labelled lectins to distinguish dysplasia from normal tissue when sprayed on to the luminal surface epithelium of freshly resected colon tissue from the Apcmin mouse and when applied to fixed human colorectal tissue sections. Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients. Changes in WGA binding to the human surface epithelium allowed regions containing normal epithelium (NE) or hyperplastic polyps (HP) to be distinguished from regions containing low-grade dysplasia (LGD), high-grade dysplasia (HGD) or carcinoma (C), with 81% sensitivity, 87% specificity and 93% positive predictive value (PPV). Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV. The decreased binding of WGA and HPA to the luminal surface epithelium in human dysplasia suggests that these lectins may enable more sensitive detection of disease in the clinic using fluorescence colonoscopy. PMID:27071814

  14. Anti-Neuroblastoma Properties of a Recombinant Sunflower Lectin

    PubMed Central

    Pinedo, Marcela; Genoula, Melanie; Silveyra, María Ximena; De Oliveira Carvalho, André; Regente, Mariana; Del Río, Marianela; Ribeiro Soares, Júlia; Moreira Gomes, Valdirene; De La Canal, Laura

    2017-01-01

    According to their sugar recognition specificity, plant lectins are proposed as bioactive proteins with potential in cancer treatment and diagnosis. Helja is a mannose-specific jacalin-like lectin from sunflower which was shown to inhibit the growth of certain fungi. Here, we report its recombinant expression in a prokaryotic system and its activity in neurobalstoma cells. Helja coding sequence was fused to the pET-32 EK/LIC, the enterokinase/Ligation-independent cloning vector and a 35 kDa protein was obtained in Escherichia coli representing Helja coupled to thioredoxin (Trx). The identity of this protein was verified using anti-Helja antibodies. This chimera, named Trx-rHelja, was enriched in the soluble bacterial extracts and was purified using Ni+2-Sepharose and d-mannose-agarose chromatography. Trx-rHelja and the enterokinase-released recombinant Helja (rHelja) both displayed toxicity on human SH-SY5Y neuroblastomas. rHelja decreased the viability of these tumor cells by 75% according to the tetrazolium reduction assay, and microscopic analyses revealed that the cell morphology was disturbed. Thus, the stellate cells of the monolayer became spheroids and were isolated. Our results indicate that rHelja is a promising tool for the development of diagnostic or therapeutic methods for neuroblastoma cells, the most common solid tumors in childhood. PMID:28075401

  15. Lectin-mediated drug delivery: the second generation of bioadhesives.

    PubMed

    Lehr, C M

    2000-03-01

    This paper reviews some recent developments in the area of bioadhesive drug delivery systems. The area of bioadhesion in drug delivery had started some 20 years ago by using so-called mucoadhesive polymers. Many of these polymers were already used as excipients in pharmaceutical formulations. This has facilitated the development of the first bioadhesive drug products, which are now commercially available. A major disadvantage of the hitherto known mucoadhesives, however, is their non-specificity with respect to the substrate. In particular for gastro-intestinal applications, this may cause some premature inactivation and moreover limits the duration of mucoadhesive bonds to the relatively fast mucus turnover. Nevertheless, for some mucoadhesive polymers other interesting functionalities were discovered, such as their ability to modulate epithelial permeability and to inhibit proteolytic enzymes. In contrast to the mucoadhesive polymers, lectins and some other adhesion molecules specifically recognize receptor-like structures of the cell membrane and therefore bind directly to the epithelial cells themselves ("cytoadhesion") rather than to the mucus gel layer. Furthermore, when bioadhesion is receptor-mediated, it is not only restricted to mere binding, but may subsequently trigger the active transport of large molecules or nanoscalic drug carrier systems by vesicular transport processes (endo-/transcytosis). Rather than only acting as a platform for controlled release systems, the concept of lectin-mediated bioadhesion therefore bears the potential for the controlled delivery of macromolecular biopharmaceuticals at relevant biological barriers, such as the epithelia of the intestinal or respiratory tract.

  16. Detection of colorectal dysplasia using fluorescently labelled lectins.

    PubMed

    Kuo, Joe Chin-Hun; Ibrahim, Ashraf E K; Dawson, Sarah; Parashar, Deepak; Howat, William J; Guttula, Kiran; Miller, Richard; Fearnhead, Nicola S; Winton, Douglas J; Neves, André A; Brindle, Kevin M

    2016-04-13

    Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions. We tested here the ability of fluorescently labelled lectins to distinguish dysplasia from normal tissue when sprayed on to the luminal surface epithelium of freshly resected colon tissue from the Apc(min) mouse and when applied to fixed human colorectal tissue sections. Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients. Changes in WGA binding to the human surface epithelium allowed regions containing normal epithelium (NE) or hyperplastic polyps (HP) to be distinguished from regions containing low-grade dysplasia (LGD), high-grade dysplasia (HGD) or carcinoma (C), with 81% sensitivity, 87% specificity and 93% positive predictive value (PPV). Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV. The decreased binding of WGA and HPA to the luminal surface epithelium in human dysplasia suggests that these lectins may enable more sensitive detection of disease in the clinic using fluorescence colonoscopy.

  17. Molecular cloning of a human macrophage lectin specific for galactose

    SciTech Connect

    Cherayil, B.J.; Chairovitz, S.; Wong, C.; Pillai, S. Harvard Medical School, Boston )

    1990-09-01

    The murine Mac-2 protein is a galactose- and IgE-binding lectin secreted by inflammatory macrophages. The authors describe here the cloning an dcharacterization of cDNA representing the human homolog of Mac-2 (hMac-2). The amino acid sequence derived from the hMac-2 cDNA indicates that the protein is evolutionarily highly conserved, with 85% of its amino acid residues being similar to those in the murine homolog. This conservation is especially marked in the carboxyl-terminal lectin domain. The amino-terminal half of the protein is less conserved but still contains the repetitive proline-glycine-rich motif seen in the mouse protein. hMac-2 synthesized in vitro is recognized by the M3/38 monoclonal antibody to Mac-2 and binds to the desialylated glycoprotein asialofetuin and to laminin, a major component of basement membranes. These findings are discussed in the context of the potential functions of hMac-2.

  18. Crystallization and crystal manipulation of the Pterocarpus angolensis seed lectin.

    PubMed

    Loris, Remy; Garcia-Pino, Abel; Buts, Lieven; Bouckaert, Julie; Beeckmans, Sonia; De Greve, Henri; Wyns, Lode

    2005-06-01

    The Man/Glc-specific legume lectin from the seeds of the African bloodwood tree (Pterocarpus angolensis) was crystallized in the presence of the disaccharide ligand Man(alpha1-3)ManMe. Small crystals initially appeared from a preliminary screen, but proved difficult to reproduce. The initial crystals were used to prepare microseeds, leading to a reproducible crystallization protocol. All attempts to obtain crystals directly of the ligand-free protein or of other carbohydrate complexes failed. However, the Man(alpha1-3)ManMe co-crystals withstand soaking with ten other carbohydrates known to bind to the lectin. Soaking for 15 min in 100 mM carbohydrate typically resulted in complete replacement of Man(alpha1-3)ManMe by the desired carbohydrate despite the involvement of lattice contacts at the binding site. Transferring the crystals for two weeks in carbohydrate-free artificial mother liquor resulted in the complete removal of the sugar from one of the two monomers in the asymmetric unit. Additional treatment of these crystals with 100 mM EDTA for two weeks resulted in removal of the structural calcium and manganese ions, which is accompanied by significant structural rearrangements of the loops that constitute the carbohydrate-binding site.

  19. Altered binding of Ulex europaeus I lectin to psoriatic epidermis.

    PubMed

    Kariniemi, A L; Holthöfer, H; Miettinen, A; Virtanen, I

    1983-11-01

    We have used Ulex europaeus I (UEA I) lectin, specific for alpha-L-fucose-containing glycoconjugates, in fluorescence microscopy to stain cryostat sections of human skin from normal persons and patients with psoriasis and lichen simplex. In normal skin the upper layers of the stratum spinosum and the stratum granulosum were strongly reactive with UEA I, whereas the lower layers of the epidermis did not react. The staining intensity of the upper epidermis was similar to that of the endothelium of dermal blood vessels. Biopsies of the lesional skin of lichen simplex showed an intense UEA I-specific staining throughout the whole epidermis, similar in intensity to that seen in the upper epidermis of normal skin. In psoriatic lesions positive UEA I-specific fluorescence was seen throughout the whole epidermis, but the fluorescence was more faint and often granular. In uninvolved skin of psoriatic patients the whole epidermis showed a diffuse UEA I-specific fluorescence, differing in this respect from normal skin. In normal skin UEA I binds to epidermal cells which are at a certain state of differentiation. The results with psoriatic epidermis confirm that both uninvolved and lesional epidermis have a defect in epidermal maturation, as shown by the altered binding of UEA I lectin.

  20. Isolation and characterization of two Korean mistletoe lectins.

    PubMed

    Kang, Tae Bong; Song, Seong Kyu; Yoon, Taek Joon; Yoo, Yung Choon; Lee, Kwan Hee; Her, Erk; Kim, Jong Bae

    2007-11-30

    Two isolectins (KML-IIU and the KML-IIL) were individually isolated from the previously reported Korean mistletoe lectin, KML-C, by using an immunoaffinity column. Molecular weights of the KML-IIU and the KML-IIL were 64 kDa and 60 kDa respectively. Both of the lectins were composed of heterogeneous A and B subunits linked with a disulfide bond, and showed the same carbohydrate-binding specificities for Gal and GalNAc. However, they are different not only in biophysical properties (glycosylation and amino acid compositions) but also bioactivities (cell killing and cytokine induction). The KML-IIL showed 17-145 times stronger in cytotoxicities to various human and mouse cancer cell lines than the KML-IIU. The KML-IIL also induced TNF-alpha secretion from mouse peritoneal macrophages 4.5 times better than the KML-IIU. The results demonstrated isolectins in Korean mistletoe were varied in bioactivities and the KML-IIL may be developed as an anti-cancer agent.

  1. Use of lectin-functionalized particles for oral immunotherapy

    PubMed Central

    Diesner, Susanne C; Wang, Xue-Yan; Jensen-Jarolim, Erika; Untersmayr, Eva; Gabor, Franz

    2013-01-01

    Immunotherapy, in recent times, has found its application in a variety of immunologically mediated diseases. Oral immunotherapy may not only increase patient compliance but may, in particular, also induce both systemic as well as mucosal immune responses, due to mucosal application of active agents. To improve the bioavailability and to trigger strong immunological responses, recent research projects focused on the encapsulation of drugs and antigens into polymer particles. These particles protect the loaded antigen from the harsh conditions in the GI tract. Furthermore, modification of the surface of particles by the use of lectins, such as Aleuria aurantia lectin, wheatgerm agglutinin or Ulex europaeus-I, enhances the binding to epithelial cells, in particular to membranous cells, of the mucosa-associated lymphoid tissue. Membranous cell-specific targeting leads to an improved transepithelial transport of the particle carriers. Thus, enhanced uptake and presentation of the encapsulated antigen by antigen-presenting cells favor strong systemic, but also local, mucosal immune responses. PMID:22834202

  2. Characterization of Caenorhabditis Elegans Lectin-Binding Mutants

    PubMed Central

    Link, C. D.; Silverman, M. A.; Breen, M.; Watt, K. E.; Dames, S. A.

    1992-01-01

    We have identified 45 mutants of Caenorhabditis elegans that show ectopic surface binding of the lectins wheat germ agglutinin (WGA) and soybean agglutinin (SBA). These mutations are all recessive and define six genes: srf-2, srf-3, srf-4, srf-5, srf-8 and srf-9. Mutations in these genes fall into two phenotypic classes: srf-2, -3, -5 mutants are grossly wild-type, except for their lectin-binding phenotype; srf-4, -8, -9 mutants have a suite of defects, including uncoordinated movement, abnormal egg laying, and defective copulatory bursae morphogenesis. Characterization of these pleiotropic mutants at the cellular level reveals defects in the migration of the gonadal distal tip cell and in axon morphology. Unexpectedly, the pleiotropic mutations also interact with mutations in the lin-12 gene, which encodes a putative cell surface receptor involved in the control of cell fate. We propose that the underlying defect in the pleiotropic mutations may be in the general processing or secretion of extracellular proteins. PMID:1516818

  3. Regulation of Adherence and Virulence by the Entamoeba histolytica Lectin Cytoplasmic Domain, Which Contains a β2 Integrin Motif

    PubMed Central

    Vines, Richard R.; Ramakrishnan, Girija; Rogers, Joshua B.; Lockhart, Lauren A.; Mann, Barbara J.; Petri, William A.

    1998-01-01

    Killing of human cells by the parasite Entamoeba histolytica requires adherence via an amebic cell surface lectin. Lectin activity in the parasite is regulated by inside-out signaling. The lectin cytoplasmic domain has sequence identity with a region of the β2 integrin cytoplasmic tail implicated in regulation of integrin-mediated adhesion. Intracellular expression of a fusion protein containing the cytoplasmic domain of the lectin has a dominant negative effect on extracellular lectin-mediated cell adherence. Mutation of the integrin-like sequence abrogates the dominant negative effect. Amebae expressing the dominant negative mutant are less virulent in an animal model of amebiasis. These results suggest that inside-out signaling via the lectin cytoplasmic domain may control the extracellular adhesive activity of the amebic lectin and provide in vivo demonstration of the lectin’s role in virulence. PMID:9693367

  4. Effective inhibition of melanosome transfer to keratinocytes by lectins and niacinamide is reversible.

    PubMed

    Greatens, Amanda; Hakozaki, Tomohiro; Koshoffer, Amy; Epstein, Howard; Schwemberger, Sandy; Babcock, George; Bissett, Donald; Takiwaki, Hirotsugu; Arase, Seiji; Wickett, R Randall; Boissy, Raymond E

    2005-07-01

    Skin pigmentation results in part from the transfer of melanized melanosomes synthesized by melanocytes to neighboring keratinocytes. Plasma membrane lectins and their glycoconjugates expressed by these epidermal cells are critical molecules involved in this transfer process. In addition, the derivative of vitamin B(3), niacinamide, can inhibit melanosome transfer and induce skin lightening. We investigated the effects of these molecules on the viability of melanocytes and keratinocytes and on the reversibility of melanosome-transfer inhibition induced by these agents using an in vitro melanocyte-keratinocyte coculture model system. While lectins and neoglycoproteins could induce apoptosis in a dose-dependent manner to melanocytes or keratinocytes in monoculture, similar dosages of the lectins, as opposed to neoglycoproteins, did not induce apoptosis to either cell type when treated in coculture. The dosages of lectins and niacinamide not affecting cell viability produced an inhibitory effect on melanosome transfer, when used either alone or together in cocultures of melanocytes-keratinocytes. Cocultures treated with lectins or niacinamide resumed normal melanosome transfer in 3 days after removal of the inhibitor, while cocultures treated with a combination of lectins and niacinamide demonstrated a lag in this recovery. Subsequently, we assessed the effect of niacinamide on facial hyperpigmented spots using a vehicle-controlled, split-faced design human clinical trial. Topical application of niacinamide resulted in a dose-dependent and reversible reduction in hyperpigmented lesions. These results suggest that lectins and niacinamide at concentrations that do not affect cell viability are reversible inhibitors of melanosome transfer.

  5. Lectin-Mediated Resistance Impairs Plant Virus Infection at the Cellular Level[C][W][OA

    PubMed Central

    Yamaji, Yasuyuki; Maejima, Kensaku; Komatsu, Ken; Shiraishi, Takuya; Okano, Yukari; Himeno, Misako; Sugawara, Kyoko; Neriya, Yutaro; Minato, Nami; Miura, Chihiro; Hashimoto, Masayoshi; Namba, Shigetou

    2012-01-01

    Plants possess a multilayered defense response, known as plant innate immunity, to infection by a wide variety of pathogens. Lectins, sugar binding proteins, play essential roles in the innate immunity of animal cells, but the role of lectins in plant defense is not clear. This study analyzed the resistance of certain Arabidopsis thaliana ecotypes to a potexvirus, plantago asiatica mosaic virus (PlAMV). Map-based positional cloning revealed that the lectin gene JACALIN-TYPE LECTIN REQUIRED FOR POTEXVIRUS RESISTANCE1 (JAX1) is responsible for the resistance. JAX1-mediated resistance did not show the properties of conventional resistance (R) protein–mediated resistance and was independent of plant defense hormone signaling. Heterologous expression of JAX1 in Nicotiana benthamiana showed that JAX1 interferes with infection by other tested potexviruses but not with plant viruses from different genera, indicating the broad but specific resistance to potexviruses conferred by JAX1. In contrast with the lectin gene RESTRICTED TEV MOVEMENT1, which inhibits the systemic movement of potyviruses, which are distantly related to potexviruses, JAX1 impairs the accumulation of PlAMV RNA at the cellular level. The existence of lectin genes that show a variety of levels of virus resistance, their targets, and their properties, which are distinct from those of known R genes, suggests the generality of lectin-mediated resistance in plant innate immunity. PMID:22307853

  6. A novel mitogenic and antiproliferative lectin from a wild cobra lily, Arisaema flavum.

    PubMed

    Singh, Jagmohan; Singh, Jatinder; Kamboj, Sukhdev Singh

    2004-06-11

    A novel lectin having specificity towards a complex glycoprotein asialofetuin was purified from tubers of Arisaema flavum (Schott.) by affinity chromatography on asialofetuin-linked amino-activated silica beads. A. flavum gave a single peak on HPLC size exclusion and a single band on non-denatured PAGE at pH 4.5. The molecular mass of the lectin, as determined by gel filtration chromatography, was 56 kDa. In SDS-PAGE, pH 8.3, the lectin migrated as a single band of 13.5 kDa, under reducing and non-reducing conditions, indicating the homotetrameric nature. A. flavum lectin (AFL) readily agglutinated rabbit, rat, sheep, goat, and guinea pig erythrocytes but not human ABO blood group erythrocytes even after neuraminidase treatment. This lectin is stable up to 55 degrees C and does not require metal ions for its hemagglutination activity. AFL was completely devoid of sulphur containing amino acids and was rich in aspartic acid and glycine. In Oucterlony's double immunodiffusion, the antisera raised against A. flavum lectin showed distinct lines of identity with those of other araceous lectins. AFL showed potent mitogenic activity towards BALB/c splenocytes and human lymphocytes in comparison to Con A, a well-known plant mitogen. AFL also showed significant in vitro antiproliferative activity towards J774 and P388D1 murine cancer cell lines.

  7. Chemical modification studies on a lectin from Saccharomyces cerevisiae (baker's yeast).

    PubMed Central

    Kundu, M; Basu, J; Ghosh, A; Chakrabarti, P

    1987-01-01

    The effect of chemical modification on a galactose-specific lectin isolated from a fatty acid auxotroph of Saccharomyces cerevisiae was investigated in order to identify the type of amino acids involved in its agglutinating activity. Modification of 50 free amino groups with succinic anhydride or citraconic anhydride led to an almost complete loss of activity. This could not be protected by the inhibitory sugar methyl alpha-D-galactopyranoside. Treatment with N-bromosuccinimide and N-acetylimidazole, for the modification of tryptophan and tyrosine residues, did not affect lectin activity. Modification of carboxy groups with glycine ethyl ester greatly affected lectin activity, although sugars afford partial protection. Modification of four thiol groups with N-ethylmaleimide was accompanied by a loss of 85% of the agglutinating activity, and two thiol groups were found to be present at the sugar-binding site of the lectin. Modification of 18 arginine residues with cyclohexane-1,2-dione and 26 histidine residues with ethoxyformic anhydride led to a loss of lectin activity. However, in these cases, modification was not protected by the abovementioned inhibitory sugar, suggesting the absence of these groups at the sugar-binding site. In all the cases, immunodiffusion studies with modified lectin showed no gross structural changes which could disrupt antigenic sites of the lectin. Images Fig. 5. PMID:3128265

  8. Lectin-Like Molecules of Lactobacillus rhamnosus GG Inhibit Pathogenic Escherichia coli and Salmonella Biofilm Formation.

    PubMed

    Petrova, Mariya I; Imholz, Nicole C E; Verhoeven, Tine L A; Balzarini, Jan; Van Damme, Els J M; Schols, Dominique; Vanderleyden, Jos; Lebeer, Sarah

    2016-01-01

    Increased antibiotic resistance has catalyzed the research on new antibacterial molecules and alternative strategies, such as the application of beneficial bacteria. Since lectin molecules have unique sugar-recognizing capacities, and pathogens are often decorated with sugars that affect their survival and infectivity, we explored whether lectins from the probiotic strain Lactobacillus rhamnosus GG have antipathogenic properties. The genome sequence of L. rhamnosus GG was screened for the presence of lectin-like proteins. Two genes, LGG_RS02780 and LGG_RS02750, encoding for polypeptides with an N-terminal conserved L-type lectin domain were detected and designated Llp1 (lectin-like protein 1) and Llp2. The capacity of Llp1 and Llp2 to inhibit biofilm formation of various pathogens was investigated. Sugar specificity was determined by Sepharose beads assays and glycan array screening. The isolated lectin domains of Llp1 and Llp2 possess pronounced inhibitory activity against biofilm formation by various pathogens, including clinical Salmonella species and uropathogenic E. coli, with Llp2 being more active than Llp1. In addition, sugar binding assays with Llp1 and Llp2 indicate specificity for complex glycans. Both proteins are also involved in the adhesion capacity of L. rhamnosus GG to gastrointestinal and vaginal epithelial cells. Lectins isolated from or expressed by beneficial lactobacilli could be considered promising bio-active ingredients for improved prophylaxis of urogenital and gastrointestinal infections.

  9. Purification and Characterization of a Lectin from Green Split Peas (Pisum sativum).

    PubMed

    Ng, Tzi Bun; Chan, Yau Sang; Ng, Charlene Cheuk Wing; Wong, Jack Ho

    2015-11-01

    Lectins have captured the attention of a large number of researchers on account of their various exploitable activities, including antitumor, immunomodulatory, antifungal, as well as HIV reverse transcriptase inhibitory activities. A mannose/glucose-specific lectin was isolated from green split peas (a variety of Pisum sativum) and characterized. The purification step involved anion-exchange chromatography on a DEAE-cellulose column, cation-exchange chromatography on an SP-Sepharose column, and gel filtration by fast protein liquid chromatography (FPLC) on Superdex 200. The purified lectin had a native molecular mass of around 50 kDa as determined by size exclusion chromatography. It appeared as a heterotetramer, composed of two distinct polypeptide bands with a molecular mass of 6 and 19 kDa, respectively, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The N-terminal sequence of green split pea lectin shows some degree of homology compared to lectins from other legume species. Its hemagglutinating activity was inhibited by glucose, mannose, and sucrose, and attenuated at pH values higher than 12 or lower than 3. Hemagglutinating activity was preserved at temperatures lower than 80 °C. The lectin did not show antifungal activity toward fungi including Fusarium oxysporum, Botrytis cinerea, and Mycosphaerella arachidicola. Green split pea lectin showed a mitogenic effect toward murine splenocytes and could inhibit the activity of HIV-1 reverse transcriptase.

  10. A novel L-fucose-binding lectin from Fenneropenaeus indicus induced cytotoxicity in breast cancer cells.

    PubMed

    Chatterjee, Biji; Ghosh, Krishna; Yadav, Nitin; Kanade, Santosh R

    2017-01-01

    Lectins are omnipresent in almost all life forms, being the proteins which specifically bind to carbohydrate moieties on the cell surface; they have been explored for their anti-tumour activities. In this study, we purified a fucose specific-lectin (IFL) from Fenneropenaeus indicus haemolymph using fucose-affinity column and characterized for its haemagglutination activity, carbohydrate specificity, dependency on cations and cytotoxicity against cancer cells. The lectin showed non-specificity against human erythrocytes. It was a Ca(2+)-dependent lectin which remained stable over wide pH and temperature ranges. The lectin showed effective dose dependent cytotoxicity against different human cancer cell lines and induced apoptosis in MCF-7 cells as evidenced by DNA ladder assay and PARP cleavage in a dose dependent manner. Moreover, an increased p21 level corresponding to cyclin D downregulation in response to IFL treatment was observed which might work as probable factors to inhibit cell growth and induce apoptosis of MCF-7 cells. Therefore, we report a novel lectin from the prawn haemolymph with high specificity for L-fucose and antiproliferative towards human cancer cells. However, further establishment of the modus operandi of this lectin is required to enable its biotechnological applications.

  11. Synthesis of a selective inhibitor of a fucose binding bacterial lectin from Burkholderia ambifaria.

    PubMed

    Richichi, Barbara; Imberty, Anne; Gillon, Emilie; Bosco, Rosa; Sutkeviciute, Ieva; Fieschi, Franck; Nativi, Cristina

    2013-06-28

    Burkholderia ambifaria is a bacterium member of the Burkholderia cepacia complex (BCC), a closely related group of Gram-negative bacteria responsible for "cepacia syndrome" in immunocompromised patients. B. ambifaria produces BambL, a fucose-binding lectin that displays fine specificity to human fucosylated epitopes. Here, we report the first example of a synthetic ligand able to selectively bind, in the micromolar range, the pathogen-lectin BambL. The synthetic routes for the preparation of the α conformationally constrained fucoside are described, focusing on a totally diastereoselective inverse electron demand [4 + 2] Diels-Alder reaction. Isothermal titration calorimetry (ITC) demonstrated that this compound binds to the pathogen-associated lectin BambL with an affinity comparable to that of natural fucose-containing oligosaccharides. No binding was observed by LecB, a fucose-binding lectin from Pseudomonas aeruginosa, and the differences in affinity between the two lectins could be rationalized by modeling. Furthermore, SPR analyses showed that this fucomimetic does not bind to the human fucose-binding lectin DC-SIGN, thus supporting the selective binding profile towards B. ambifaria lectin.

  12. BEL β-trefoil: a novel lectin with antineoplastic properties in king bolete (Boletus edulis) mushrooms.

    PubMed

    Bovi, Michele; Cenci, Lucia; Perduca, Massimiliano; Capaldi, Stefano; Carrizo, Maria E; Civiero, Laura; Chiarelli, Laurent R; Galliano, Monica; Monaco, Hugo L

    2013-05-01

    A novel lectin was purified from the fruiting bodies of king bolete mushrooms (Boletus edulis, also called porcino, cep or penny bun). The lectin was structurally characterized i.e its amino acid sequence and three-dimensional structure were determined. The new protein is a homodimer and each protomer folds as β-trefoil domain and therefore we propose the name Boletus edulis lectin (BEL) β-trefoil to distinguish it from the other lectin that has been described in these mushrooms. The lectin has potent anti-proliferative effects on human cancer cells, which confers to it an interesting therapeutic potential as an antineoplastic agent. Several crystal forms of the apoprotein and of complexes with different carbohydrates were studied by X-ray diffraction. The structure of the apoprotein was solved at 1.12 Å resolution. The interaction of the lectin with lactose, galactose, N-acetylgalactosamine and T-antigen disaccharide, Galβ1-3GalNAc, was examined in detail. All the three potential binding sites present in the β-trefoil fold are occupied in at least one crystal form and are described in detail in this paper. No important conformational changes are observed in the lectin when comparing its co-crystals with carbohydrates with those of the ligand-free protein.

  13. Comparative Study of Lectin Domains in Model Species: New Insights into Evolutionary Dynamics

    PubMed Central

    Van Holle, Sofie; De Schutter, Kristof; Eggermont, Lore; Tsaneva, Mariya; Dang, Liuyi; Van Damme, Els J. M.

    2017-01-01

    Lectins are present throughout the plant kingdom and are reported to be involved in diverse biological processes. In this study, we provide a comparative analysis of the lectin families from model species in a phylogenetic framework. The analysis focuses on the different plant lectin domains identified in five representative core angiosperm genomes (Arabidopsis thaliana, Glycine max, Cucumis sativus, Oryza sativa ssp. japonica and Oryza sativa ssp. indica). The genomes were screened for genes encoding lectin domains using a combination of Basic Local Alignment Search Tool (BLAST), hidden Markov models, and InterProScan analysis. Additionally, phylogenetic relationships were investigated by constructing maximum likelihood phylogenetic trees. The results demonstrate that the majority of the lectin families are present in each of the species under study. Domain organization analysis showed that most identified proteins are multi-domain proteins, owing to the modular rearrangement of protein domains during evolution. Most of these multi-domain proteins are widespread, while others display a lineage-specific distribution. Furthermore, the phylogenetic analyses reveal that some lectin families evolved to be similar to the phylogeny of the plant species, while others share a closer evolutionary history based on the corresponding protein domain architecture. Our results yield insights into the evolutionary relationships and functional divergence of plant lectins. PMID:28587095

  14. Lectin-Like Molecules of Lactobacillus rhamnosus GG Inhibit Pathogenic Escherichia coli and Salmonella Biofilm Formation

    PubMed Central

    Petrova, Mariya I.; Imholz, Nicole C. E.; Verhoeven, Tine L. A.; Balzarini, Jan; Van Damme, Els J. M.; Schols, Dominique; Vanderleyden, Jos; Lebeer, Sarah

    2016-01-01

    Objectives Increased antibiotic resistance has catalyzed the research on new antibacterial molecules and alternative strategies, such as the application of beneficial bacteria. Since lectin molecules have unique sugar-recognizing capacities, and pathogens are often decorated with sugars that affect their survival and infectivity, we explored whether lectins from the probiotic strain Lactobacillus rhamnosus GG have antipathogenic properties. Methods The genome sequence of L. rhamnosus GG was screened for the presence of lectin-like proteins. Two genes, LGG_RS02780 and LGG_RS02750, encoding for polypeptides with an N-terminal conserved L-type lectin domain were detected and designated Llp1 (lectin-like protein 1) and Llp2. The capacity of Llp1 and Llp2 to inhibit biofilm formation of various pathogens was investigated. Sugar specificity was determined by Sepharose beads assays and glycan array screening. Results The isolated lectin domains of Llp1 and Llp2 possess pronounced inhibitory activity against biofilm formation by various pathogens, including clinical Salmonella species and uropathogenic E. coli, with Llp2 being more active than Llp1. In addition, sugar binding assays with Llp1 and Llp2 indicate specificity for complex glycans. Both proteins are also involved in the adhesion capacity of L. rhamnosus GG to gastrointestinal and vaginal epithelial cells. Conclusions Lectins isolated from or expressed by beneficial lactobacilli could be considered promising bio-active ingredients for improved prophylaxis of urogenital and gastrointestinal infections. PMID:27537843

  15. Quantitation of two endogenous lactose-inhibitable lectins in embryonic and adult chicken tissues

    SciTech Connect

    Beyer, E.C.; Barondes, S.H.

    1982-01-01

    Two lactose-binding lectins from chicken tissues, chicken-lactose-lectin-I (CLL-I) and chicken-lactose-lectin-II (CLL-II) were quantified with a radioimmunoassay in extracts of a number of developing and adult chicken tissues. Both lectins could be measured in the same extract without separation, because they showed no significant immunological cross- reactivity. Many embryonic and adult tissues, including brain, heart, intestine, kidney, liver, lung, muscle, pancreas, and spleen, contained one or both lectins, although their concentrations differed markedly. For example, embryonic muscle, the richest source of CLL-I contained only traces of CLL-II whereas embryonic kidney, a very rich source of CLL-II contained substantial CLL-I. In both muscle and kidney, lectin levels in adulthood were much lower than in the embryonic state. In contrast, CLL-I in liver and CLL-II in intestine were 10-fold to 30-fold more concentrated in the adult than in the 15-d embryo. CLL-I and CLL-II from several tissues were purified by affinity chromatography and their identity in the various tissues was confirmed by polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping. The results suggest that these lectins might have different functions in the many developing and adult tissues in which they are found.

  16. Proteins with an Euonymus lectin-like domain are ubiquitous in Embryophyta

    PubMed Central

    2009-01-01

    Background Cloning of the Euonymus lectin led to the discovery of a novel domain that also occurs in some stress-induced plant proteins. The distribution and the diversity of proteins with an Euonymus lectin (EUL) domain were investigated using detailed analysis of sequences in publicly accessible genome and transcriptome databases. Results Comprehensive in silico analyses indicate that the recently identified Euonymus europaeus lectin domain represents a conserved structural unit of a novel family of putative carbohydrate-binding proteins, which will further be referred to as the Euonymus lectin (EUL) family. The EUL domain is widespread among plants. Analysis of retrieved sequences revealed that some sequences consist of a single EUL domain linked to an unrelated N-terminal domain whereas others comprise two in tandem arrayed EUL domains. A new classification system for these lectins is proposed based on the overall domain architecture. Evolutionary relationships among the sequences with EUL domains are discussed. Conclusion The identification of the EUL family provides the first evidence for the occurrence in terrestrial plants of a highly conserved plant specific domain. The widespread distribution of the EUL domain strikingly contrasts the more limited or even narrow distribution of most other lectin domains found in plants. The apparent omnipresence of the EUL domain is indicative for a universal role of this lectin domain in plants. Although there is unambiguous evidence that several EUL domains possess carbohydrate-binding activity further research is required to corroborate the carbohydrate-binding properties of different members of the EUL family. PMID:19930663

  17. Isolation and analysis of mannose/trehalose/maltose specific lectin from jack bean with antibruchid activity.

    PubMed

    Shanmugavel, Sakthivelkumar; Velayutham, Veeramani; Kamalanathan, Tamilarasan; Periasamy, Mullainadhan; Munusamy, Arumugam; Sundaram, Janarthanan

    2016-10-01

    A lectin with insecticidal property against the stored product pest, Callosobruchus maculatus was successfully isolated from the seeds of Canavalia virosa using standard affinity chromatography. The isolated molecule typically behaved like a lectin in its characteristics. It agglutinated indicator red blood cells (RBC) in its native as well as enzyme treated conditions. The enzyme treated RBC types exhibited a very high hemagglutination (HA) titre values and this property of isolated molecule behaved like arcelin, the lectin-like molecules reported from several species of Phaseolus. As a characteristic feature of a lectin, the isolated molecule effectively inhibited the agglutination of indicator RBC types with simple and complex carbohydrates including glycoproteins. This nature of the isolated molecule also relate with characteristic feature of arcelin isoforms in inhibiting HA activity with complex glycoproteins as reported in many studies. Most interestingly, the present study disclosed trehalose as a potent inhibitor of C. virosa lectin. Therefore, feeding insect pests on the lectin like arcelin could serve as antibiosis factor/anti-insect activity. The molecular characteristics of this isolated molecule and its mass studies too revealed its homology with arcelin, arcelin-1, 2 and 6 isoforms of P. vulgaris and lectin from Canavalia cathartica, C. lineata and C. brasiliensis.

  18. Characterization and molecular cloning of mannose-binding lectins from the Orchidaceae species Listera ovata, Epipactis helleborine and Cymbidium hybrid.

    PubMed

    Van Damme, J M; Smeets, K; Torrekens, S; Van Leuven, F; Peumans, W J

    1994-04-15

    Mannose-binding lectins were purified from the leaves of three Orchidaceae species, namely Listera ovata (twayblade), Epipactis helleborine (broad-leaved helleborine) and Cymbidium hybrid, using affinity chromatography on Mannose - Sepharose-4B. Apparently, the Orchidaceae lectins are dimeric proteins composed of lectin subunits of 12-13 kDa. All of the isolated lectins exhibit exclusive specificity towards mannose. A cDNA library constructed from poly(A) rich RNA isolated from leaves of L. ovata was screened for cDNA clones encoding the lectin using colony hybridization. Since N-terminal sequence analysis of the twayblade lectin revealed some sequence similarity to the previously cloned mannose-binding lectin Hippeastrum hybrid (amaryllis) ovaries, the amaryllis lectin cDNA clone was used as a probe to screen the L. ovata library. Subsequently, the cDNA clone encoding the L. ovata lectin was used to screen the cDNA libraries from the taxonomically related orchid species Cymbidium hybrid and E. helleborine. Sequence analysis of the lectin cDNA clones from different Orchidaceae species revealed approximately 50% sequence similarity both at the nucleotide and amino acid level. The Orchidaceae lectins are apparently translated from mRNAs consisting of approximately 800 nucleotides. The primary translation products are preproproteins which are converted into the mature lectins following post-translational modifications. Southern blot analysis of genomic DNA has shown that the lectins are most probably encoded by a family of closely related genes which is in good agreement with the sequence heterogeneity found between different lectin cDNA clones of one species.

  19. Thermodynamic and kinetic analysis of carbohydrate binding to the basic lectin from winged bean (Psophocarpus tetragonolobus).

    PubMed

    Khan, M I; Sastry, M V; Surolia, A

    1986-03-05

    A basic lectin (pI approximately 10.0) was purified to homogeneity from the seeds of winged bean (Psophocarpus tetragonolobus) by affinity chromatography on Sepharose 6-aminocaproyl-D-galactosamine. The lectin agglutinated trypsinized rabbit erythrocytes and had a relative molecular mass of 58,000 consisting of two subunits of Mr 29,000. The lectin binds to N-dansylgalactosamine, leading to a 15-fold increase in dansyl fluorescence with a concomitant 25-nm blue shift in the emission maximum. The lectin has two binding sites/dimer for this sugar and an association constant of 4.17 X 10(5) M-1 at 25 degrees C. The strong binding to N-dansylgalactosamine is due to a relatively positive entropic contribution as revealed by the thermodynamic parameters: delta H = -33.62 kJ mol-1 and delta S0 = -5.24 J mol-1 K-1. Binding of this sugar to the lectin shows that it can accommodate a large hydrophobic substituent on the C-2 carbon of D-galactose. Studies with other sugars indicate that a hydrophobic substituent in alpha-conformation at the anomeric position increases the affinity of binding. The C-4 and C-6 hydroxyl groups are critical for sugar binding to this lectin. Lectin difference absorption spectra in the presence of N-acetylgalactosamine indicate perturbation of tryptophan residues on sugar binding. The results of stopped flow kinetics with N-dansylgalactosamine and the lectin are consistent with a simple one-step mechanism for which k+1 = 1.33 X 10(4) M-1 s-1 and k-1 = 3.2 X 10(-2) s-1 at 25 degrees C. This k-1 is slower than any reported for a lectin-monosaccharide complex so far. The activation parameters indicate an enthalpically controlled association process.

  20. Lectin-based immunoassay for aberrant IgG glycosylation as the biomarker for Crohn's disease.

    PubMed

    Shinzaki, Shinichiro; Kuroki, Eri; Iijima, Hideki; Tatsunaka, Norika; Ishii, Mayuko; Fujii, Hironobu; Kamada, Yoshihiro; Kobayashi, Taku; Shibukawa, Narihiro; Inoue, Takahiro; Tsujii, Masahiko; Takeishi, Shunsaku; Mizushima, Tsunekazu; Ogata, Atsushi; Naka, Tetsuji; Plevy, Scott E; Takehara, Tetsuo; Miyoshi, Eiji

    2013-02-01

    Easily measured and clinically useful biomarkers for inflammatory bowel disease (IBD) are required to advance patient care. We previously reported that the agalactosyl fraction among fucosylated IgG oligosaccharides is increased in IBD, especially Crohn's disease (CD). The present study aimed to establish a simple detection system for aberrant glycosylated IgG based on lectin-oligosaccharide interactions. Lectins with higher affinity to serum IgG from IBD patients than healthy volunteers (HV) were screened by lectin microarray. Binding of selected lectins to agalactosyl IgG was definitively confirmed using step-by-step glycosidase treatment. Using the selected lectins, a lectin-enzyme-linked immunosorbent assay system was established and its clinical utility was investigated in a total of 410 (249 Japanese and 161 American) IBD patients, disease controls, and HVs. Agaricus bisporus Agglutinin (ABA) and Griffonia simplicifolia Lectin-II (GSL-II) had higher affinity for serum agalactosyl IgG from IBD patients, especially those with CD, compared to HV. Agalactosyl IgG levels measured by a lectin-enzyme immunoassay (EIA) with ABA or GSL-II were significantly increased in CD compared with HV and disease controls. Agalactosyl IgG levels significantly correlated with disease activity, showed higher predictability of therapeutic outcomes for CD than C-reactive protein levels, and exhibited higher specificity for diagnosing IBD in combination with anti-Saccharomyces cerevisiae antibody (ASCA). Validation analysis showed that agalactosyl IgG levels were significantly increased in Japanese and American CD patients. A lectin-EIA for agalactosyl IgG is a novel biomarker for IBD, especially in patients with CD.

  1. Physicochemical properties and oxidative inactivation of soluble lectin from water buffalo (Bubalus bubalis) brain.

    PubMed

    Rizvi, Sabika; Banu, Naheed

    2008-03-01

    Lectins are carbohydrate-binding proteins present in a wide variety of plants and animals, which serve various important physiological functions. A soluble beta-galactoside binding lectin has been isolated and purified to homogeneity from buffalo brain using ammonium sulphate precipitation (40-70%) and gel permeation chromatography on Sephadex G50-80 column. The molecular weight of buffalo brain lectin (BBL) as determined by SDS-PAGE under reducing and non-reducing conditions was 14.2 kDa, however, with gel filtration it was 28.5 kDa, revealing the dimeric form of protein. The neutral sugar content of the soluble lectin was estimated to be 3.3%. The BBL showed highest affinity for lactose and other sugar moieties in glycosidic form, suggesting it to be a beta-galactoside binding lectin. The association constant for lactose binding as evidenced by Scatchard analysis was 6.6 x 10(3) M(-1) showing two carbohydrate binding sites per lectin molecule. A total inhibition of lectin activity was observed by denaturants like guanidine HCl, thiourea and urea at 6 M concentration. The treatment of BBL with oxidizing agent destroyed its agglutination activity, abolished its fluorescence, and shifted its UV absorption maxima from 282 to 250 nm. The effect of H2O2 was greatly prevented by lactose indicating that BBL is more stable in the presence of its specific ligand. The purified lectin was investigated for its brain cell aggregation properties by testing its ability to agglutinate cells isolated from buffalo and goat brains. Rate of aggregation of buffalo brain cells by purified protein was more than the goat brain cells. The data from above study suggests that the isolated lectin may belong to the galectin-1 family but is glycosylated unlike those purified till date.

  2. Putative glycoprotein and glycolipid polymorphonuclear leukocyte receptors for the Actinomyces naeslundii WVU45 fimbrial lectin.

    PubMed Central

    Sandberg, A L; Ruhl, S; Joralmon, R A; Brennan, M J; Sutphin, M J; Cisar, J O

    1995-01-01

    Recognition of receptors on sialidase-treated polymorphonuclear leukocytes (PMNs) by the Gal/GalNAc lectin associated with the type 2 fimbriae of certain strains of actinomyces results in activation of the PMNs, phagocytosis, and destruction of the bacteria. In the present study, plant lectins were utilized as probes to identify putative PMN receptors for the actinomyces lectin. The Gal-reactive lectin from Ricinus communis (RCAI), the Gal/GalNAc-reactive lectins from R. communis (RCAII) and Bauhinia purpurea (BPA), as well as the Gal beta 1-3GalNAc-specific lectins from Arachis hypogaea (PNA) and Agaricus bisporus (ABA) inhibited killing of Actinomyces naeslundii WVU45 by sialidase-treated PMNs. These five lectins detected a 130-kDa surface-labeled glycoprotein on nitrocellulose transfers of PMN extracts separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This glycoprotein was revealed only after treatment of the transfers with sialidase, a condition analogous to the sialidase dependence of the lectin-mediated biological responses of the PMNs to the actinomyces. The mannose-reactive lectin concanavalin A did not inhibit killing of the actinomyces and failed to detect the 130-kDa glycoprotein but did block PMN-dependent killing of Escherichia coli B, a bacterium that possesses mannose-sensitive fimbriae. Therefore, the PMN glycoprotein receptor for A. naeslundii is clearly distinct from those recognized by E. coli. Two major putative glycolipid receptors were also identified by actinomyces and RCAI overlays on sialidase-treated thin-layer chromatograms of PMN gangliosides. Thus, both a 130-kDa glycoprotein and certain gangliosides are implicated in the attachment of the actinomyces to PMNs. PMID:7790078

  3. Receptor mobility and the binding of cells to lectin-coated fibers

    PubMed Central

    1975-01-01

    The ability of cells to bind to nylon fibers coated with lectin molecules interspaced with varying numbers of albumin molecules has been analyzed. The cells used were lymphoma cells, normal lymphocytes, myeloid leukemia cells, and normal and transformed fibroblasts, and the fibers were coated with different densities of concanavalin A or the lectins from soybean or wheat germ. Cells fixed with glutaraldehyde did not bind to lectin-coated fibers. The number of cells bound to fibers could be increased by increasing the density of lectin molecules on the fiber, the density of specific receptors on the cell, or the mobility of the receptors. It is suggested that binding of cells to fibers involves alignment and binding of specific cell surface receptors with lectin molecules immobilized on the fibers, and that this alignment requires short-range rapid lateral mobility (RLM) of the receptors. The titration of cell binding to fibers coated with different densities of lectin and albumin has been used to measure the relative RLM of unoccupied cell surface receptors for the lectin. The results indicate a relationship of RLM to lectin-induced cell-to-cell binding. The RLM or receptors for concanavalin A (Con A) was generally found to be higher than that of receptors for the lectins from wheat germ or soybean. Receptor RLM could be decreased by use of metabolic inhibitors or by lowering the temperature. Receptors for Con A had a lower RLM on normal fibroblasts than on SV40-transformed fibroblasts, and trypsinization of normal fibroblasts increased Con A receptor RLM. Normal lymphocytes, lymphoma cells, and lines of myeloid leukemia cells that can be induced to differentiate had a high receptor RLM, whereas lines of myeloid leukemia cells that could not be induced to differentiate had a low receptor RLM. These results suggest that the RLM of Con A receptors is related to the transformation of fibroblasts and the ability of myeloid leukemia cells to undergo differentiation PMID

  4. Differentiation of Bacillus Anthracis and Other Bacillus Species by Use of Lectins

    DTIC Science & Technology

    1983-07-18

    TITL9 fAnd Subtfitle) S.TypeO REPORT gi PZRCC rvt 4 DIFFERENTIATION OF BACIL-LUSg’ ANTHRAtgACIS D OTHER BACILLUS , SPECIES BY-USE OYLECTINS" Inter[im...Ricinus communis. Some strains of Bacillus cer-eus var. m-ycoides (B. Mycoides) were strongly reactive with the lectin from Helbi pomtia and weakly reacti...ve with the Glycine max lectin. The differential iCnteractions between Bacillus species and lectins af forded a means of distinguishing B. anthracis

  5. Artocarpin is a polyspecific jacalin-related lectin with a monosaccharide preference for mannose.

    PubMed

    Barre, Annick; Peumans, Willy J; Rossignol, Michel; Borderies, Gisèle; Culerrier, Raphaël; Van Damme, Els J M; Rougé, Pierre

    2004-01-01

    A reinvestigation of the carbohydrate-binding properties revealed that artocarpin, a previously described mannose-specific lectin from jackfruit (Artocarpus integrifolia) seeds, behaves as a polyspecific lectin. Surface plasmon resonance hapten inhibition experiments demonstrated that artocarpin readily interacted with a wide range of monosaccharides covering galactose, N-acetylgalactosamine, mannose, glucose, sialic acid and N-acetylmuramic acid. Molecular docking confirmed this unexpected ability of artocarpin to interact with structurally different sugars. The biological significance of the polyspecificity of the lectin is discussed in terms of the broadening of the range of potential target glycans present on the surface of plant phytopathogens or predators.

  6. Biomimics of fungal cell-cell recognition by use of lectin-coated nylon fibers.

    PubMed Central

    Inbar, J; Chet, I

    1992-01-01

    When the mycoparasitic, biocontrol fungus Trichoderma harzianum was allowed to grow on nylon fibers treated with concanavalin A or Sclerotium rolfsii lectin, it coiled around the nylon fibers and produced hooks in a pattern similar to that observed with the real host hyphae. The incidence of interaction between T. harzianum and S. rolfsii lectin-treated fibers was significantly higher than that of the controls (untreated or blocked activated fibers). These findings provide direct evidence for the role of lectins in mycoparasitism. Images PMID:1732197

  7. Sugar recognition and protein-protein interaction of mammalian lectins conferring diverse functions.

    PubMed

    Nagae, Masamichi; Yamaguchi, Yoshiki

    2015-10-01

    Recent advances in structural analyses of mammalian lectins reveal atomic-level details of their fine specificities toward diverse endogenous and exogenous glycans. Local variations on a common scaffold can enable certain lectins to recognize complex carbohydrate ligands including branched glycans and O-glycosylated peptides. Simultaneous recognition of both glycan and the aglycon moieties enhances the affinity and specificity of lectins such as CLEC-2 and PILRα. Attention has been paid to the roles of galectin and RegIII family of proteins in protein-protein interactions involved in critical biological functions including signal transduction and bactericidal pore formation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Genomic sequence and organization of two members of a human lectin gene family

    SciTech Connect

    Gitt, M.A.; Barondes, S.H. )

    1991-01-01

    The authors have isolated and sequenced the genomic DNA encoding a human dimeric soluble lactose-binding lectin. The gene has four exons, and its upstream region contains sequences that suggest control by glucocorticoids, heat (environmental) shock, metals, and other factors. They have also isolated and sequenced three exons of the gene encoding another human putative lectin, the existence of which was first indicated by isolation of its cDNA. Comparisons suggest a general pattern of genomic organization of members of this lectin gene family.

  9. Influence of glycosidic linkage on the nature of carbohydrate binding in beta-prism I fold lectins: an X-ray and molecular dynamics investigation on banana lectin-carbohydrate complexes.

    PubMed

    Sharma, Alok; Vijayan, Mamannamana

    2011-01-01

    The three crystal structures reported here provide details of the interactions of mannose and the mannosyl-α-1,3-mannose component of a pentamannose with banana lectin and evidence for the binding of glucosyl-α-1,2-glucose to the lectin. The known structures involving the lectin include a complex with glucosyl-β-1,3-glucose. Modeling studies on the three disaccharide complexes with the reducing end and the nonreducing end at the primary binding site are also provided here. The results of the X-ray and modeling studies show that the disaccharides with an α-1,3 linkage prefer to have the nonreducing end at the primary binding site, whereas the reducing end is preferred at the site when the linkage is β-1,3 in mannose/glucose-specific β-prism I fold lectins. In the corresponding galactose-specific lectins, however, α-1,3-linked disaccharides cannot bind the lectin with the nonreducing end at the primary binding site on account of steric clashes with an aromatic residue that occurs only when the lectin is galactose-specific. Molecular dynamics simulations based on the known structures involving banana lectin enrich the information on lectin-carbohydrate interactions obtained from crystal structures. They demonstrate that conformational selection as well as induced fit operate when carbohydrates bind to banana lectin.

  10. Structural and functional diversity of the lectin repertoire in teleost fish: Relevance to innate and adaptive immunity

    PubMed Central

    Vasta, Gerardo R.; Nita-Lazar, Mihai; Giomarelli, Barbara; Ahmed, Hafiz; Du, Shaojun; Cammarata, Matteo; Parrinello, Nicolò; Bianchet, Mario A.; Amzel, L. Mario

    2012-01-01

    Protein–carbohydrate interactions mediated by lectins have been recognized as key components of innate immunity in vertebrates and invertebrates, not only for recognition of potential pathogens, but also for participating in downstream effector functions, such as their agglutination, immobilization, and complement-mediated opsonization and killing. More recently, lectins have been identified as critical regulators of mammalian adaptive immune responses. Fish are endowed with virtually all components of the mammalian adaptive immunity, and are equipped with a complex lectin repertoire. In this review, we discuss evidence suggesting that: (a) lectin repertoires in teleost fish are highly diversified, and include not only representatives of the lectin families described in mammals, but also members of lectin families described for the first time in fish species; (b) the tissue-specific expression and localization of the diverse lectin repertoires and their molecular partners is consistent with their distinct biological roles in innate and adaptive immunity; (c) although some lectins may bind endogenous ligands, others bind sugars on the surface of potential pathogens; (d) in addition to pathogen recognition and opsonization, some lectins display additional effector roles, such as complement activation and regulation of immune functions; (e) some lectins that recognize exogenous ligands mediate processes unrelated to immunity: they may act as anti-freeze proteins or prevent polyspermia during fertilization. PMID:21896283

  11. Cloning of cDNAs encoding C-type lectins from Elapidae snakes Bungarus fasciatus and Bungarus multicinctus.

    PubMed

    Zha, H G; Lee, W H; Zhang, Y

    2001-12-01

    A number of C-type lectins with various biological activities have been purified and characterized from Viperidae snake venoms. In contrast, only a few reports could be found in literature concerning the C-type lectins in Elapidae snake venoms. Based on the published cDNA sequences of C-type lectins from Viperidae snake venoms, oligonucleotide primers were designed and used to screen the cDNA libraries made from the venom glands of Bungarus fasciatus and Bungarus multicinctus. This allowed the cloning of three full length cDNAs encoding C-type lectins. The encoded proteins, named BFL-1, BFL-2 and BML, exhibit high degrees of sequence identities with Viperidae snake venom saccharide-binding lectins (around 60% with Trimeresurus stejnegeri venom lectin, Crotalus atrox venom lectin and Agkistrodon piscivorus venom lectin). They show much less identities with other venom C-type lectin-like proteins (around 30% with the platelet glycoprotein Ib-binding protein from Agkistrodon blomhoffi venom and the factor IX/X-binding protein from Trimeresurus flavoviridis venom). The cDNAs revealed that the precursors contain potential signal peptides characterized by a hydrophobic core. To our knowledge, these are the first cDNA cloning of group VII C-type lectins (Drickamer K. 1993. Prog. Nucleic Acid Res. Mol. Biol. 45, 207-232) from Elapidae snake venom glands.

  12. Differential Lectin Binding Patterns Identify Distinct Heart Regions in Giant Danio ( Devario aequipinnatus) and Zebrafish ( Danio rerio) Hearts.

    PubMed

    Manalo, Trina; May, Adam; Quinn, Joshua; Lafontant, Dominique S; Shifatu, Olubusola; He, Wei; Gonzalez-Rosa, Juan M; Burns, Geoffrey C; Burns, Caroline E; Burns, Alan R; Lafontant, Pascal J

    2016-11-01

    Lectins are carbohydrate-binding proteins commonly used as biochemical and histochemical tools to study glycoconjugate (glycoproteins, glycolipids) expression patterns in cells, tissues, including mammalian hearts. However, lectins have received little attention in zebrafish ( Danio rerio) and giant danio ( Devario aequipinnatus) heart studies. Here, we sought to determine the binding patterns of six commonly used lectins-wheat germ agglutinin (WGA), Ulex europaeus agglutinin, Bandeiraea simplicifolia lectin (BS lectin), concanavalin A (Con A), Ricinus communis agglutinin I (RCA I), and Lycopersicon esculentum agglutinin (tomato lectin)-in these hearts. Con A showed broad staining in the myocardium. WGA stained cardiac myocyte borders, with binding markedly stronger in the compact heart and bulbus. BS lectin, which stained giant danio coronaries, was used to measure vascular reconstruction during regeneration. However, BS lectin reacted poorly in zebrafish. RCA I stained the compact heart of both fish. Tomato lectin stained the giant danio, and while low reactivity was seen in the zebrafish ventricle, staining was observed in their transitional cardiac myocytes. In addition, we observed unique staining patterns in the developing zebrafish heart. Lectins' ability to reveal differential glycoconjugate expression in giant danio and zebrafish hearts suggests they can serve as simple but important tools in studies of developing, adult, and regenerating fish hearts.

  13. Cell- and region-specific expression of sugar chains in the mouse epididymal epithelium using lectin histochemistry combined with immunohistochemistry.

    PubMed

    Tajiri, Shota; Fukui, Tatsuya; Sawaguchi, Akira; Yoshinaga, Kazuya

    2012-02-01

    To understand the cytochemical properties of epididymal epithelial cells, the characteristics of glycoconjugates in the mouse epididymis were examined using the technique of lectin histochemistry combined with immunohistochemistry. Characteristic staining patterns depending on the type of lectins were observed in the epididymal epithelium. Principal cells expressed N-acetyl-D-galactosamine (GalNAc), N-acetyl-D-glucosamine (GlcNAc), and Fucose in the proximal region of the epididymis and Mannose, Glucose, and Galactose in the distal region of the epididymis. Basal cells expressed Mannose, Glucose, Galactose, and GlcNAc in the proximal region and Galactose in the distal region. On the other hand, clear cells expressed various sugar residues and differences among regions were not observed. Interestingly, principal cells, clear cells, and basal cells specifically reacted with Ulex Europaeus-Agglutinin I (UEA-I lectin), Maackia Amurensis-Lectin I (MAL-I lectin), and Griffonia simplicifolia Lectin I-B4 (GS-I B lectin), respectively. These findings indicate that the selectivity in lectin reactivity for distinct cell types and segment-dependent staining in the epididymis may be related to cellular and regional differences in function. Furthermore, because some lectins stain particular cells or cellular compartments selectively, these lectins could be useful markers for histopathological evaluation of diseases or diagnosis of male infertility.

  14. A novel L-type lectin was required for the multiplication of WSSV in red swamp crayfish (Procambarus clakii).

    PubMed

    Dai, Yunjia; Wang, Yuqing; Zhao, Lingling; Qin, Zhendong; Yuan, Junfa; Qin, Qiwei; Lin, Li; Lan, Jiangfeng

    2016-08-01

    L-type lectins are involved in glycoproteins secretory pathways and are associated with many immune responses. There is growing evidence that L-type lectins are also involved in viral replication. In this study, a novel L-type lectin (named as PcL-lectin) was identified from red swamp crayfish (Procambarus clakii). Gene sequencing and phylogenetic tree analysis results showed that the PcL-lectin was a kind of endoplasmic reticulum Golgi intermediate compartment-53 (ERGIC-53). The expression level of PcL-lectin was significantly down regulated in crayfish after challenged with white spot syndrome virus (WSSV). Recombinant PcL-lectin protein facilitated the replication of WSSV in crayfish. In addition, WSSV replication was decreased when endogenous PcL-lectin was knocked down by RNA interference in crayfish. Furthermore, PcL-lectin may interact with VP24, an envelope protein of WSSV. Our results suggest that PcL-lectin may be required for the multiplication of WSSV, and will pave a new way for the developing of strategies against WSSV infection.

  15. A comparative study of lectin affinity based plant N-glycoproteome profiling using tomato fruit as a model.

    PubMed

    Ruiz-May, Eliel; Hucko, Simon; Howe, Kevin J; Zhang, Sheng; Sherwood, Robert W; Thannhauser, Theodore W; Rose, Jocelyn K C

    2014-02-01

    Lectin affinity chromatography (LAC) can provide a valuable front-end enrichment strategy for the study of N-glycoproteins and has been used to characterize a broad range eukaryotic N-glycoproteomes. Moreover, studies with mammalian systems have suggested that the use of multiple lectins with different affinities can be particularly effective. A multi-lectin approach has also been reported to provide a significant benefit for the analysis of plant N-glycoproteins; however, it has yet to be determined whether certain lectins, or combinations of lectins are optimal for plant N-glycoproteome profiling; or whether specific lectins show preferential association with particular N-glycosylation sites or N-glycan structures. We describe here a comparative study of three mannose-binding lectins, concanavalin A, snowdrop lectin, and lentil lectin, to profile the N-glycoproteome of mature green stage tomato (Solanum lycopersicum) fruit pericarp. Through coupling lectin affinity chromatography with a shotgun proteomics strategy, we identified 448 putative N-glycoproteins, whereas a parallel lectin affinity chromatography plus hydrophilic interaction chromatography analysis revealed 318 putative N-glycosylation sites on 230 N-glycoproteins, of which 100 overlapped with the shotgun analysis, as well as 17 N-glycan structures. The use of multiple lectins substantially increased N-glycoproteome coverage and although there were no discernible differences in the structures of N-glycans, or the charge, isoelectric point (pI) or hydrophobicity of the glycopeptides that differentially bound to each lectin, differences were observed in the amino acid frequency at the -1 and +1 subsites of the N-glycosylation sites. We also demonstrated an alternative and complementary in planta recombinant expression strategy, followed by affinity MS analysis, to identify the putative N-glycan structures of glycoproteins whose abundance is too low to be readily determined by a shotgun approach, and

  16. Atomic- Resolution Crystal Structure of the Antiviral Lectin Scytovirin

    SciTech Connect

    Moulaei,T.; Botos, I.; Ziolkowska, N.; Bokesch, H.; Krumpe, L.; McKee, T.; O'Keefe, B.; Dauter, Z.; Wlodawer, A.

    2007-01-01

    The crystal structures of the natural and recombinant antiviral lectin scytovirin (SVN) were solved by single-wavelength anomalous scattering and refined with data extending to 1.3 Angstroms and 1.0 Angstroms resolution, respectively. A molecule of SVN consists of a single chain 95 amino acids long, with an almost perfect sequence repeat that creates two very similar domains (RMS deviation 0.25 Angstroms for 40 pairs of Ca atoms). The crystal structure differs significantly from a previously published NMR structure of the same protein, with the RMS deviations calculated separately for the N- and C-terminal domains of 5.3 Angstroms and 3.7 Angstroms, respectively, and a very different relationship between the two domains. In addition, the disulfide bonding pattern of the crystal structures differs from that described in the previously published mass spectrometry and NMR studies.

  17. Data on IL-17 production induced by plant lectins

    PubMed Central

    da Silva, Thiago Aparecido; Fernandes, Fabrício Freitas; Roque-Barreira, Maria Cristina

    2016-01-01

    We reported in article da Silva et al. (2016) [2] that ArtinM induces the IL-17 production through interaction with CD4+ T cells and stimulation of IL-23 and IL-1. Besides ArtinM, other plant lectins (PLs) induce IL-17 production by murine spleen cells. The IL-17 production induced by PLs was evaluated regarding the involvement of IL-23, IL-6, Th1-, and Th2-cytokines. Furthermore, the effect exerted TLR2, TLR4, and CD14 on the PLs׳ performance in the induction of IL-17 was examined. The current data were compared to the known ArtinM ability to induce Th17 immunity. PMID:27222857

  18. Biodiversity of mannose-specific lectins within Narcissus species.

    PubMed

    Lopez, Susanna; Codina, Carlos; Bastida, Jaume; Viladomat, Francesc; Davidson, Elaine; Stewart, Derek

    2002-04-24

    Mannose-specific lectins (MSLs) were isolated from the bulbs of 27 species of wild Spanish Narcissi and compared to the commercially available MSL from daffodil (Narcissus pseudonarcissus, NPA). Molecular weight analysis showed the monomers of all the MSLs were at, or around, 12.5 kD. Haemagglutination assays showed that the MSLs exhibited activities at up to four times greater than that displayed by NPA and other MSLs derived from other species such as Galanthus nivalus (snowdrop) and Allium ursinum (ramson). Elution profiles from ion exchange chromatography exhibited similarities for species within the same taxonomic section suggesting that this method could aid in species classification. Further analysis by isoelectric focusing showed many isolectins are present in vivo and that even within a single peak from ion exchange chromatography there are numerous isolectins present. The basis of the isolectin heterogeneity is suggested to reside in the tetraploidy (sometime triploidy) nature of Narcissus genes.

  19. Direct Measurement of Glyconanoparticles and Lectin Interactions by Isothermal Titration Calorimetry

    PubMed Central

    Wang, Xin; Matei, Elena; Gronenborn, Angela M.; Ramström, Olof; Yan, Mingdi

    2012-01-01

    Glyconanomaterials have shown high potential in applications including bioanalysis and nanomedicine. Here, a quantitative analytical technique, based on isothermal titration calorimetry, was developed to characterize the interactions between glyconanoparticles and lectins. By titrating lectins into the glyconanoparticle solution, the apparent dissociation constant, thermodynamic parameters, and the number of binding sites were derived simultaneously. For the glyconanoparticles-lectin binding pairs investigated, a 3–5 orders of magnitude affinity enhancement over the free ligand-lectin interactions was observed which can be attributed to the multivalent ligand presentation on the nanoparticles. The impact of ligand density was also studied, and results showed that the affinity increased with the number of glycans on the nanoparticle. PMID:22548468

  20. Direct measurement of glyconanoparticles and lectin interactions by isothermal titration calorimetry.

    PubMed

    Wang, Xin; Matei, Elena; Gronenborn, Angela M; Ramström, Olof; Yan, Mingdi

    2012-05-15

    Glyconanomaterials have shown high potential in applications including bioanalysis and nanomedicine. Here, a quantitative analytical technique, based on isothermal titration calorimetry, was developed to characterize the interactions between glyconanoparticles and lectins. By titrating lectins into the glyconanoparticle solution, the apparent dissociation constant, thermodynamic parameters, and the number of binding sites were derived simultaneously. For the glyconanoparticles-lectin binding pairs investigated, a 3-5 order of magnitude affinity enhancement over the free ligand-lectin interactions was observed which can be attributed to the multivalent ligand presentation on the nanoparticles. The impact of ligand density was also studied, and results showed that the affinity increased with the number of glycans on the nanoparticle.

  1. Novel hemagglutinating, hemolytic and cytotoxic activities of the intermediate subunit of Entamoeba histolytica lectin

    PubMed Central

    Kato, Kentaro; Yahata, Kazuhide; Gopal Dhoubhadel, Bhim; Fujii, Yoshito; Tachibana, Hiroshi

    2015-01-01

    Galactose and N-acetyl-D-galactosamine (Gal/GalNAc) inhibitable lectin of Entamoeba histolytica, a common protozoan parasite, has roles in pathogenicity and induction of protective immunity in mouse models of amoebiasis. The lectin consists of heavy (Hgl), light (Lgl), and intermediate (Igl) subunits. Hgl has lectin activity and Lgl does not, but little is known about the activity of Igl. In this study, we assessed various regions of Igl for hemagglutinating activity using recombinant proteins expressed in Escherichia coli. We identified a weak hemagglutinating activity of the protein. Furthermore, we found novel hemolytic and cytotoxic activities of the lectin, which resided in the carboxy-terminal region of the protein. Antibodies against Igl inhibited the hemolytic activity of Entamoeba histolytica trophozoites. This is the first report showing hemagglutinating, hemolytic and cytotoxic activities of an amoebic molecule, Igl. PMID:26354528

  2. pH and glucose responsive nanofibers for the reversible capture and release of lectins.

    PubMed

    Wang, Yinan; Kotsuchibashi, Yohei; Uto, Koichiro; Ebara, Mitsuhiro; Aoyagi, Takao; Liu, Yang; Narain, Ravin

    2015-01-01

    A dual pH and glucose responsive boronic acid containing nanofiber was constructed for the reversible capture and release of lectins. The effects of surface groups and pH values on selective lectin capture were investigated by fluorescence microscopy. Compared to the pristine nanofibrous membrane, glucose and galactose functionalized nanofiber surfaces showed significantly higher capture of ConA and Jacalin, under alkaline conditions. On the other hand, treatment of the modified nanofibers with an acidic solution resulted in the detachment of both the lectins and glycopolymers from the nanofiber surface. As expected, once the glycopolymers are displaced, no lectins were adhered to the nanofiber surface under alkaline conditions. These functional nanofibers can therefore be easily modified and hence can be used for quick removal of selective proteins or toxins from the solution.

  3. Entomotoxic and nematotoxic lectins and protease inhibitors from fungal fruiting bodies.

    PubMed

    Sabotič, Jerica; Ohm, Robin A; Künzler, Markus

    2016-01-01

    Fruiting bodies or sporocarps of dikaryotic (ascomycetous and basidiomycetous) fungi, commonly referred to as mushrooms, are often rich in entomotoxic and nematotoxic proteins that include lectins and protease inhibitors. These protein toxins are thought to act as effectors of an innate defense system of mushrooms against animal predators including fungivorous insects and nematodes. In this review, we summarize current knowledge about the structures, target molecules, and regulation of the biosynthesis of the best characterized representatives of these fungal defense proteins, including galectins, beta-trefoil-type lectins, actinoporin-type lectins, beta-propeller-type lectins and beta-trefoil-type chimerolectins, as well as mycospin and mycocypin families of protease inhibitors. We also present an overview of the phylogenetic distribution of these proteins among a selection of fungal genomes and draw some conclusions about their evolution and physiological function. Finally, we present an outlook for future research directions in this field and their potential applications in medicine and crop protection.

  4. Novel hemagglutinating, hemolytic and cytotoxic activities of the intermediate subunit of Entamoeba histolytica lectin.

    PubMed

    Kato, Kentaro; Yahata, Kazuhide; Gopal Dhoubhadel, Bhim; Fujii, Yoshito; Tachibana, Hiroshi

    2015-09-10

    Galactose and N-acetyl-D-galactosamine (Gal/GalNAc) inhibitable lectin of Entamoeba histolytica, a common protozoan parasite, has roles in pathogenicity and induction of protective immunity in mouse models of amoebiasis. The lectin consists of heavy (Hgl), light (Lgl), and intermediate (Igl) subunits. Hgl has lectin activity and Lgl does not, but little is known about the activity of Igl. In this study, we assessed various regions of Igl for hemagglutinating activity using recombinant proteins expressed in Escherichia coli. We identified a weak hemagglutinating activity of the protein. Furthermore, we found novel hemolytic and cytotoxic activities of the lectin, which resided in the carboxy-terminal region of the protein. Antibodies against Igl inhibited the hemolytic activity of Entamoeba histolytica trophozoites. This is the first report showing hemagglutinating, hemolytic and cytotoxic activities of an amoebic molecule, Igl.

  5. Secretory Expression and Characterization of Chinese Narcissus GNA-Like Lectin in Pichia pastoris.

    PubMed

    Li, Xinyu; Zhang, Pengpeng; Zhang, Ning; Liang, Xiao; Wang, Wenya; Yuan, Qipeng; Li, Qiang

    2017-08-01

    Narcissus tazetta lectin (NTL) is a GNA-like lectin, which has a wide potential application in medicine, agriculture, and glycobiology. In the present paper, the codon-optimized ntl gene was transformed into the yeast Pichia pastoris; SDS-PAGE gel and western blotting analysis revealed that the recombinant lectin was expressed successfully in Pichia yeast. The similarity between the recombinant NTL and the native NTL was confirmed by circular dichroism (CD) and hemagglutination assay further. In the 5-L scale fermentator, the protein yield was as high as 1.2 g/L after fermentation for 96 h. In addition, the effect of metal ions (K(+), Mg(2+), Ca(2+), and Cu(2+)), acid, and alkaline on hemagglutinating activity of NTL was tested, which provided biochemical characterizations of the mannose-binding lectin from Chinese Narcissus.

  6. [Dynamic of lectin activity during germination of bean seeds (Phaseolus vulgaris L.)].

    PubMed

    Koval'chuk, N V

    2006-01-01

    PHA quantity and activity dynamics during early germination of bean seed were investigated. Electrophoretic characteristics, subunits composition and carbohydrate-binding specificity of lectin extracted from white kidney bean cv. Bilozerna were studied. It was shown that investigated lectin consisted of 2 subunits E and L with molecular weight 34 and 36 kDa, respectively, analogously to purified PHA ("Serva", Germany), and specifically bound N-acetyl-D-galactosamin and galactose. During germination both quantity and activity of PHA were dramatically decreasing in embryonic axes and in cotyledons, possibly, as a result of the lectin release from seeds to the environment. It is very likely that one of the defence mechanisms of germinating seeds is related with the releasing of lectins that are able to bind components of the bacterial cell wall and to inhibit their growth.

  7. Lectin-Magnetic Beads for Plasma Membrane Isolation.

    PubMed

    Lee, Yu-Chen; Liu, Hsuan-Chen; Chuang, Carol; Lin, Sue-Hwa

    2015-07-01

    Plasma membrane proteins mainly function to transmit external signals into the cell. Many plasma membrane receptor tyrosine kinases (e.g., HER2 and EGFR) are known to mediate oncogenic progression, making them prime targets for cancer therapy. Recently, it has become important to identify plasma membrane proteins that are differentially expressed in normal versus cancer cells, in drug-sensitive versus drug-resistant cells, or among tumor cells that metastasize to different organ sites because these differentially expressed membrane proteins may lead to the identification of therapeutic targets or diagnostic markers. In addition, there is an increased interest in identifying cell-surface proteins that could serve as markers for stem cells, progenitor cells, or cells of different lineages. Traditionally, membrane isolation requires multiple centrifugation steps to isolate different organelles based on their density. With the advent of affinity matrix technology, it is possible to separate organelles based on their molecular differences. A defining characteristic of the plasma membrane is that plasma membrane proteins are more extensively glycosylated than are intracellular membrane proteins. As a result, affinity chromatography employing lectin, a carbohydrate-binding protein, is commonly used to isolate plasma membrane proteins. We have extended this concept for plasma membrane isolation by using concanavalin A (ConA), a lectin with mannose specificity. Here we describe a protocol that uses immobilized ConA bound to magnetic beads to isolate plasma membranes from homogenized cell lysates. The captured plasma membrane proteins are then solubilized from the ConA-magnetic beads by detergents in the presence of a competing sugar, methyl α-mannopyranoside.

  8. Evidence for an increase in positive surface charge and an increase in susceptibility to trypsin of Sarcophaga lectin (from the flesh fly, Sarcophaga peregrina) on its interaction with galactose, a hapten sugar of the lectin.

    PubMed

    Komano, H; Kurama, T; Nagasawa, Y; Natori, S

    1992-05-15

    When Sarcophaga lectin (from the flesh fly, Sarcophaga peregrina), an insect humoral lectin, was eluted from a column of DEAE-cellulose in the presence of galactose (a hapten sugar of this lectin), it emerged at a lower salt concentration than when galactose was absent. In the presence of galactose the lectin was, in addition, more susceptible to trypsin digestion. The lectin was found to have an affinity for basic proteins such as histone H3 and sarcotoxin IA, but this property was lost in the presence of galactose. These results suggested that the lectin changes its conformation on interaction with galactose. This change is suggested to result in the exposure of some hidden lysine and/or arginine residues.

  9. Evidence for an increase in positive surface charge and an increase in susceptibility to trypsin of Sarcophaga lectin (from the flesh fly, Sarcophaga peregrina) on its interaction with galactose, a hapten sugar of the lectin.

    PubMed Central

    Komano, H; Kurama, T; Nagasawa, Y; Natori, S

    1992-01-01

    When Sarcophaga lectin (from the flesh fly, Sarcophaga peregrina), an insect humoral lectin, was eluted from a column of DEAE-cellulose in the presence of galactose (a hapten sugar of this lectin), it emerged at a lower salt concentration than when galactose was absent. In the presence of galactose the lectin was, in addition, more susceptible to trypsin digestion. The lectin was found to have an affinity for basic proteins such as histone H3 and sarcotoxin IA, but this property was lost in the presence of galactose. These results suggested that the lectin changes its conformation on interaction with galactose. This change is suggested to result in the exposure of some hidden lysine and/or arginine residues. Images Fig. 1. Fig. 3. PMID:1599400

  10. Integrative analysis workflow for the structural and functional classification of C-type lectins

    PubMed Central

    2011-01-01

    Background It is important to understand the roles of C-type lectins in the immune system due to their ubiquity and diverse range of functions in animal cells. It has been observed that currently confirmed C-type lectins share a highly conserved domain known as the C-type carbohydrate recognition domain (CRD). Using the sequence profile of the CRD, an increasing number of putative C-type lectins have been identified. Hence, it is highly needed to develop a systematic framework that enables us to elucidate their carbohydrate (glycan) recognition function, and discover their physiological and pathological roles. Results Presented herein is an integrated workflow for characterizing the sequence and structural features of novel C-type lectins. Our workflow utilizes web-based queries and available software suites to annotate features that can be found on the C-type lectin, given its amino acid sequence. At the same time, it incorporates modeling and analysis of glycans - a major class of ligands that interact with C-type lectins. Thereafter, the results are analyzed together with context-specific knowledge to filter off unlikely predictions. This allows researchers to design their subsequent experiments to confirm the functions of the C-type lectins in a systematic manner. Conclusions The efficacy and usefulness of our proposed immunoinformatics workflow was demonstrated by applying our integrated workflow to a novel C-type lectin -CLEC17A - and we report some of its possible functions that warrants further validation through wet-lab experiments. PMID:22372988

  11. Genome-wide analysis of lectin receptor-like kinases in Populus

    DOE PAGES

    Yang, Yongil; Labbé, Jessy; Muchero, Wellington; ...

    2016-09-01

    Receptor-like kinases (RLKs) belong to a large protein family with over 600 members in Arabidopsis and over 1000 in rice. Among RLKs, the lectin receptor-like kinases (LecRLKs) possess a characteristic extracellular carbohydrate-binding lectin domain and play important roles in plant development and innate immunity. In addition, there are 75 and 173 LecRLKs in Arabidopsis and rice, respectively. However, little is known about LecRLKs in perennial woody plants.

  12. Genome-wide analysis of lectin receptor-like kinases in Populus

    SciTech Connect

    Yang, Yongil; Labbé, Jessy; Muchero, Wellington; Yang, Xiaohan; Jawdy, Sara S.; Kennedy, Megan; Johnson, Jenifer; Sreedasyam, Avinash; Schmutz, Jeremy; Tuskan, Gerald A.; Chen, Jin -Gui

    2016-09-01

    Receptor-like kinases (RLKs) belong to a large protein family with over 600 members in Arabidopsis and over 1000 in rice. Among RLKs, the lectin receptor-like kinases (LecRLKs) possess a characteristic extracellular carbohydrate-binding lectin domain and play important roles in plant development and innate immunity. In addition, there are 75 and 173 LecRLKs in Arabidopsis and rice, respectively. However, little is known about LecRLKs in perennial woody plants.

  13. Glycoconjugates in the mandibular salivary gland of adult dogs revealed by lectin histochemistry.

    PubMed

    Pedini, V; Ceccarelli, P; Gargiulo, A M

    1994-11-01

    The glycosidic residues in the mandibular glands of five adult dogs were studied by using seven different lectin-horseradish peroxidase conjugates. In some cases a treatment with sialidase preceded the lectin staining. The mucous acinar cells contained oligosaccharides with alpha- and beta-N-acetylgalactosamine, N-acetylglucosamine and fucose residues, whereas the demilunar cells contained glycoconjugates rich in sialic acid linked to the penultimate disaccharide galactosyl-(beta 1-->3) N-acetylgalactosamine.

  14. Carbohydrate mimics and lectins: a source of new drugs and therapeutic opportunities.

    PubMed

    Reina, Jose J; Bernardi, Anna

    2012-12-01

    Mimics of oligosaccharides capable of interfering with lectin activity are currently being pursued by a number of groups in an effort to produce tools for glycobiology and to design antagonists of medically relevant lectins. The field is reviewed in this chapter. After a brief overview of the state of the art, examples from our and others' studies on the dendritic cell receptor DC-SIGN are illustrated.

  15. Fucofullerenes as tight ligands of RSL and LecB, two bacterial lectins.

    PubMed

    Buffet, Kevin; Gillon, Emilie; Holler, Michel; Nierengarten, Jean-François; Imberty, Anne; Vincent, Stéphane P

    2015-06-21

    A series of water-soluble glycofullerenes containing up to 24 fucose residues have been prepared. These compounds were tested against the two bacterial fucose-binding lectins LecB and RSL, and C60(E)12 bearing 24 fucose residues appeared to be the best known inhibitor of both lectins to date. We have shown that increasing both the valency and the length of the spacer between the central core and the peripheral sugars can be beneficial for the affinity.

  16. A C-Type Lectin from Bothrops jararacussu Venom Disrupts Staphylococcal Biofilms

    PubMed Central

    Klein, Raphael Contelli; Fabres-Klein, Mary Hellen; de Oliveira, Leandro Licursi; Feio, Renato Neves; Malouin, François; Ribon, Andréa de Oliveira Barros

    2015-01-01

    Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16) showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model. PMID:25811661

  17. Isolation and antiproliferative activity of Lotus corniculatus lectin towards human tumour cell lines.

    PubMed

    Rafiq, Shaista; Majeed, Rabiya; Qazi, Asif Khurshid; Ganai, Bashir Ahmad; Wani, Ishfak; Rakhshanda, Syed; Qurishi, Yasrib; Sharma, P R; Hamid, Abid; Masood, Akbar; Hamid, Rabia

    2013-12-15

    The objective of the study was to investigate the anti cancer activity of a lectin isolated from Lotus corniculatus seeds. A tetrameric 70kDa galactose specific lectin was purified using two step simple purification protocol which involved affinity chromatography on AF-BlueHC650M and gel filtration on Sephadex G-100. The lectin was adsorbed on AF-BlueHC650M and desorbed using 1M NaCl in the starting buffer. Gel filtration on Sephadex G-100 yielded a major peak absorbance that gave two bands of 15kDa and 20kDa in SDS PAGE. Hemagglutination activity was completely preserved, when the temperature was in the range of 20-60°C. However, drastic reduction in activity occurred at temperatures above 60°C. Full hemagglutination activity was retained at ambient pH 4-12. Thereafter no activity was observed above pH 13. Hemaglutination of the lectin was inhibited by d-galactose. The lectin showed a strong antiproliferative activity towards human leukemic (THP-1) cancer cells followed by lung cancer (HOP62) cells and HCT116 with an IC50 of 39μg/ml and 50μg/ml and 60μg/ml respectively. Flow cytometry analysis showed an increase in the percentage of cells in sub G0G1 phase confirming that Lotus corniculatus lectin induced apoptosis. Morphological observations showed that Lotus corniculatus lectin (LCL) treated THP-1 cells displayed apparent apoptosis characteristics such as nuclear fragmentation, appearance of membrane enclosed apoptotic bodies and DNA fragmentation. Lotus corniculatus lectin (LCL) effectively inhibits the cell migration in a dose dependent manner as indicated by the wound healing assay.

  18. Metal ions in sugar binding, sugar specificity and structural stability of Spatholobus parviflorus seed lectin.

    PubMed

    Abhilash, Joseph; Dileep, Kalarickal Vijayan; Palanimuthu, Muthusamy; Geethanandan, Krishnan; Sadasivan, Chittalakkotu; Haridas, Madhathilkovilakath

    2013-08-01

    Spatholobus parviflorus seed lectin (SPL) is a heterotetrameric lectin, with two α and two β monomers. In the crystal structure of SPL α monomer, two residues at positions 240 and 241 are missing. This region was modeled based on the positional and sequence similarities. The role of metal ions in SPL structure was analyzed by 10 ns molecular dynamics simulation. MD simulations were performed in the presence and absence of metal ions to explain the loss of haemagglutinating property of the lectin due to demetallization. Demetallized structure was found to deviate drastically at the metal binding loop region. Affinity of different sugars like N-acetyl galactosamine (GalNAc), D-galactose and lactose towards the native and demetallized protein was calculated by molecular docking studies. It was found that the sugar binding site got severely distorted in demetallized lectin. Consequently, sugar binding ability of lectin might be decreasing in the demetallized condition. Isothermal titration calorimetric (ITC) analysis of the sugars in the presence of native and demetallized protein confirmed the in silico results. It was observed after molecular dynamics simulations, that significant structural deviations were not caused in the quaternary structure of demetallized lectin. It was confirmed that the structural changes modified the sugar binding ability, as well as sugar specificity of the present lectin. The role of metal ions in sugar binding is described based on the in silico studies and ITC analysis. A comprehensive analysis of the ITC data suggests that the sugar specificity of the metal bound lectin and the loss of sugar specificity due to metal chelation are not linear.

  19. Seasonal Fluctuations of Lectins in Barks of Elderberry (Sambucus nigra) and Black Locust (Robinia pseudoacacia) 1

    PubMed Central

    Nsimba-Lubaki, Makuta; Peumans, Willy J.

    1986-01-01

    Elderberry (Sambucus nigra) and black locust (Robinia pseudoacacia) agglutinins, which are abundantly present in the bark of both species, display seasonal fluctuations with regard to their content in this tissue. These seasonal changes result apparently from a circa-annual rhythm of lectin accumulation and depletion during autumn and spring, respectively. Because the bark of trees can be considered as a type of vegetative storage tissue, the results suggest that bark lectins behave as typical storage proteins. Images Fig. 4 PMID:16664696

  20. Lectin staining patterns in human gastric mucosae with and without exposure to Helicobacter pylori

    PubMed Central

    Melo-Junior, Mario R.; Cavalcanti, Carmelita L.B.; Pontes-Filho, Nicodemos T.; Carvalho Jr, Luiz B.; Beltrão, Eduardo I. C.

    2008-01-01

    The aim of the present study was to evaluate qualitative changes in the glycoconjugate expression in human gastric tissue of positive and negative patients for Helicobacter pylori, through lectins: Wheat Germ Agglutinin (WGA) and Concanavalin A (Con A). The lectins recognized differently the glycoconjugates in the superficial mucous layer at the gastric tissues. The results suggest a significant change in the carbohydrate moieties present on the surface of the gastric cells during infection. PMID:24031208

  1. Potential use of the Macrobrachium rosenbergii lectin for diagnosis of T-cell acute lymphoblastic leukemia.

    PubMed

    Pérez-Campos-Mayoral, Laura; Ruiz-Argüelles, Alejandro; Pérez-Romano, Beatriz; Zenteno, Edgar; Hernández-Cruz, Pedro; Martínez-Cruz, Ruth; Martínez-Cruz, Margarito; Pina-Canseco, Socorro; Pérez-Campos, Eduardo

    2008-01-01

    T-cell acute lymphoblastic leukemia is the most common form of cancer in children. Lectins are proteins or glycoproteins from plants or animals that recognize oligossacharides on the cell surface and have been used to characterize the structural changes of oligosaccharides in leukemias. In this study, we used the lectin from the freshwater prawn Macrobrachium (M. rosenbergii), specific for acetyl groups in sialylated glycans, because increased sialylation of glycoproteins and glycolipids has been identified in lymphoblastic leukemias. We compared the specificity of the M. rosenbergii lectin for lymphoblastic leukemias with the specificities of the lectins from Triticum vulgaris, Solanum tuberosum, Arachis hipogaea, and Phytolacca americana. By morphologic and phenotype characterization with a panel of monoclonal antibodies, we identified four types of leukemias from 106 leukemia patients: 11 cases of T-cell acute lymphoblastic leukemia, 61 cases of B-cell acute lymphoblastic leukemia, 24 cases of acute myeloblastic leukemia, and 10 cases of acute biphenotypic leukemia. As determined by cytofluorometric assays, nine of the eleven cases with T-cell acute lymphoblastic leukemia (8 +/- 3 years old) were specifically identified with the lectin from M. rosenbergii. In contrast, only six cases of B-cell leukemia, one case of myeloblastic leukemia, and 2 cases of biphenotypic leukemia were identified with this M. rosenbergii lectin. The other lectins tested showed no capacity to differentiate, in a significant manner, any of the four types of leukemias tested. Thus, the lectin from M. rosenbergii could be considered a useful tool for the diagnosis and study of T-cell acute lymphoblastic leukemia.

  2. Lectin Binding to the Root and Root Hair Tips of the Tropical Legume Macroptilium atropurpureum Urb

    PubMed Central

    Ridge, R. W.; Rolfe, B. G.

    1986-01-01

    Ten fluorescein isothiocyanate-labeled lectins were tested on the roots of the tropical legume Macroptilium atropurpureum Urb. Four of these (concanavalin A, peanut agglutinin, Ricinis communis agglutinin I [RCA-I], wheat germ agglutinin) were found to bind to the exterior of root cap cells, the root cap slime, and the channels between epidermal cells in the root elongation zone. One of these lectins, RCA-I, bound to the root hair tips in the mature and emerging hair zones and also to sites at which root hairs were only just emerging. There was no RCA-I binding to immature trichoblasts. Preincubation of these lectins with their hapten sugars eliminated all types of root cell binding. By using a microinoculation technique, preincubation of the root surface with RCA-I lectin was found to inhibit infection and nodulation by Rhizobium spp. Preincubation of the root surface with the RCA-I hapten β-d-galactose or a mixture of RCA-I lectin and its hapten failed to inhibit nodulation. Application of RCA-I lectin to the root surface caused no apparent detrimental effects to the root hair cells and did not prevent the growth of root hairs. The lectin did not prevent Rhizobium sp. motility or viability even after 24 h of incubation. It was concluded that the RCA-I lectin-specific sugar β-d-galactose may be involved in the recognition or early infection stages, or both, in the Rhizobium sp. infection of M. atropurpureum. Images PMID:16346989

  3. Interactions with lectins and agglutination profiles of clinical, food, and environmental isolates of Listeria.

    PubMed Central

    Facinelli, B; Giovanetti, E; Casolari, C; Varaldo, P E

    1994-01-01

    On the basis of preliminary trials with 14 collection strains of Listeria, five lectins (Canavalia ensiformis, concanavalin A; Griffonia simplicifolia lectin I; Helix pomatia agglutinin; Ricinus communis agglutinin; and Triticum vulgaris wheat germ agglutinin) were selected to set up a microtiter agglutination assay. The lectin agglutination profiles of 174 clinical, food, and environmental strains of Listeria monocytogenes, Listeria innocua, and Listeria seeligeri were investigated. Data on the standard determination of the antigenic structure were available for clinical strains; nonclinical isolates were assigned to serogroup 1 or 4 with commercial antisera. The listeria-lectin interaction was related to serological type rather than species; in particular, the strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, 3a, 3b, and 7 were never agglutinated by G. simplicifolia lectin I. The five-lectin set proved to be capable of detecting differences between serologically identical isolates of L. monocytogenes. Of the 150 isolates of this species, 144 were distributed over 15 different lectin agglutination profiles and 6 autoagglutinated, the overall typeability being 96%. However, the profiles encountered among L. monocytogenes isolates were not randomly distributed. With strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, and 3b, the clinical isolates fell into only two of the eight patterns recorded overall; with strains of serogroup 4 and serovar 4b, food and environmental isolates were distributed over eight of the nine patterns found in total, while clinical isolates were distributed over five patterns. In a comparative study of 15 epidemiologically relevant isolates of L. monocytogenes from five distinct outbreaks, strains with identical phage types and/or DNA fingerprints displayed identical lectin profiles. The heterogeneity of agglutination profiles may form the basis of a new approach to L. monocytogenes typing

  4. Odorranalectin Is a Small Peptide Lectin with Potential for Drug Delivery and Targeting

    PubMed Central

    Xu, Xueqing; Yang, Hailong; Wu, Bingxian; Wang, Yipeng; Zhu, Jianhua; Lai, Ren; Jiang, Xinguo; Lin, Donghai; Prescott, Mark C.; Rees, Huw H.

    2008-01-01

    Background Lectins are sugar-binding proteins that specifically recognize sugar complexes. Based on the specificity of protein–sugar interactions, different lectins could be used as carrier molecules to target drugs specifically to different cells which express different glycan arrays. In spite of lectin's interesting biological potential for drug targeting and delivery, a potential disadvantage of natural lectins may be large size molecules that results in immunogenicity and toxicity. Smaller peptides which can mimic the function of lectins are promising candidates for drug targeting. Principal Findings Small peptide with lectin-like behavior was screened from amphibian skin secretions and its structure and function were studied by NMR, NMR-titration, SPR and mutant analysis. A lectin-like peptide named odorranalectin was identified from skin secretions of Odorrana grahami. It was composed of 17 aa with a sequence of YASPKCFRYPNGVLACT. L-fucose could specifically inhibit the haemagglutination induced by odorranalectin. 125I-odorranalectin was stable in mice plasma. In experimental mouse models, odorranalectin was proved to mainly conjugate to liver, spleen and lung after i.v. administration. Odorranalectin showed extremely low toxicity and immunogenicity in mice. The small size and single disulfide bridge of odorranalectin make it easy to manipulate for developing as a drug targeting system. The cyclic peptide of odorranalectin disclosed by solution NMR study adopts a β-turn conformation stabilized by one intramolecular disulfide bond between Cys6-Cys16 and three hydrogen bonds between Phe7-Ala15, Tyr9-Val13, Tyr9-Gly12. Residues K5, C6, F7, C16 and T17 consist of the binding site of L-fucose on odorranalectin determined by NMR titration and mutant analysis. The structure of odorranalectin in bound form is more stable than in free form. Conclusion These findings identify the smallest lectin so far, and show the application potential of odorranalectin for drug

  5. Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation.

    PubMed

    Biswas, Himadri; Chattopadhyaya, Rajagopal

    2016-04-01

    Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules.

  6. Inhibition of hepatitis C virus by the cyanobacterial protein Microcystis viridis lectin: mechanistic differences between the high-mannose specific lectins MVL, CV-N, and GNA.

    PubMed

    Kachko, Alla; Loesgen, Sandra; Shahzad-Ul-Hussan, Syed; Tan, Wendy; Zubkova, Iryna; Takeda, Kazuyo; Wells, Frances; Rubin, Steven; Bewley, Carole A; Major, Marian E

    2013-12-02

    Plant or microbial lectins are known to exhibit potent antiviral activities against viruses with glycosylated surface proteins, yet the mechanism(s) by which these carbohydrate-binding proteins exert their antiviral activities is not fully understood. Hepatitis C virus (HCV) is known to possess glycosylated envelope proteins (gpE1E2) and to be potently inhibited by lectins. Here, we tested in detail the antiviral properties of the newly discovered Microcystis viridis lectin (MVL) along with cyanovirin-N (CV-N) and Galanthus nivalis agglutinin (GNA) against cell culture HCV, as well as their binding properties toward viral particles, target cells, and recombinant HCV glycoproteins. Using infectivity assays, CV-N, MVL, and GNA inhibited HCV with IC50 values of 0.6 nM, 30.4 nM, and 11.1 nM, respectively. Biolayer interferometry analysis demonstrated a higher affinity of GNA to immobilized recombinant HCV glycoproteins compared to CV-N and MVL. Complementary studies, including fluorescence-activated cell sorting (FACS) analysis, confocal microscopy, and pre- and post-virus binding assays, showed a complex mechanism of inhibition for CV-N and MVL that includes both viral and cell association, while GNA functions by binding directly to the viral particle. Combinations of GNA with CV-N or MVL in HCV infection studies revealed synergistic inhibitory effects, which can be explained by different glycan recognition profiles of the mainly high-mannoside specific lectins, and supports the hypothesis that these lectins inhibit through different and complex modes of action. Our findings provide important insights into the mechanisms by which lectins inhibit HCV infection. Overall, the data suggest MVL and CV-N have the potential for toxicity due to interactions with cellular proteins while GNA may be a better therapeutic agent due to specificity for the HCV gpE1E2.

  7. Molecular cloning of a lectin cDNA from Alocasia macrorrhiza and prediction of its characteristics.

    PubMed

    Zhu, Ya-Ran; Wang, Jie; Huang, Bing-Qiu; Hou, Xue-Wen

    2006-12-01

    The cDNA of Alocasia macrorrhiza lectin (aml, GenBank accession number: DQ340864) was cloned by RACE-PCR and its characteristics were predicted by various bioinformatics tools. GSPs (Gene Specific Primers) were designed according to the conserved regions of the genes encoded for lectins and similar proteins from the same family Araceae. Total RNAs were extracted from the tubers of A macrorrhiza by Qiagen RNeasy mini kit. The 3'- and 5'-RACE-PCRs were performed with the isolated total RNAs by SMART(TM)RACE cDNA amplification kit from BD Biosciences Clontech Company, respectively. The purified PCR products were ligated with pMD 18-T vector, and the confirmed clones were sequenced. The full-length cDNA of aml was obtained by combination of 3'- and 5'-end sequences, and was then confirmed by full-length 3'-RACE-PCR. The aml cDNA is 1 124 bp long. The deduced amino acid length of AML lectin is 270 aa. Its relative molecular weight is 29.7 kD. The results of homologous analysis showed a high similarity between AML and other mannose-binding lectins and similar proteins from Araceae family. Two typical B-lectin domains and three mannose- binding motifs were found in the sequence of AML. With all these taken together, it can be concluded that this newly cloned aml cDNA encodes for a mannose-binding lectin.

  8. Interactions of Pseudomonas aeruginosa PA-IIL lectin with quail egg white glycoproteins.

    PubMed

    Lerrer, B; Gilboa-Garber, N

    2001-12-01

    Pseudomonas aeruginosa produces several lectins, including the galactophilic PA-IL and the fucose- and mannose-binding PA-IIL. The great advantage of these two lectins is their stability in purified preparations. Following observations that pigeon egg white blocks Escherichia coli P-fimbriae and PA-IL, we examined the interactions of diverse avian egg white components with PA-IIL. This lectin may represent both mannose- and fucose-specific microbial adhesins. For comparison, Con A (which also binds mannose) and Ulex europaeus lectin (UEA-I, which binds fucose) were analyzed in parallel. The lectin interactions with chicken, quail, and pigeon egg whites and several purified chicken egg white glycoproteins were examined by a hemagglutination inhibition test and Western blotting. Both analyses showed that like Con A and unlike UEA-I, which was not sensitive to any of these three egg whites, PA-IIL most strongly reacted with the quail egg white. However, in contrast with Con A, its interactions with the chicken egg white components, excluding avidin, were very poor. The results of this study might indicate the possibility that some of the egg white components that interacted with the above two mannose-binding lectins (exhibiting individual heterogeneity) might be associated with the innate immunity against mannose-specific microbial or viral adhesion during the fowl embryonic period.

  9. Influence of fetal bovine serum on cytotoxic and genotoxic effects of lectins in MCF-7 cells.

    PubMed

    Faheina-Martins, Glaucia Veríssimo; da Silveira, Alethéia Lacerda; Ramos, Márcio Viana; Marques-Santos, Luís Fernando; Araujo, Demetrius Antonio Machado

    2011-01-01

    Canavalia ensiformis (ConA), Canavalia brasiliensis (Conbr), and Cratylia floribunda (CFL) lectins have exhibited glucose-mannose binding specificity. We investigated the effect of fetal bovine serum (FBS) concentrations (1, 5, 10, and 20%) on the cytotoxic effect of these lectins against breast tumor cell line MCF-7. Cell viability was examined using the MTT reduction assay. When cells were grown in a medium supplemented with a higher serum concentration (10 or 20%), all lectins were much less toxic. When we used 1% FBS, it was possible to achieve a concentration-dependent activity by all examined lectins, with an IC(50) of 3.5, 25, and 60 µg/mL for ConA, Conbr, and CFL, respectively. All lectins incubated with 1% FBS induced apoptosis and DNA damage in MCF-7 cells. We conclude that ConA, Conbr, and CFL lectins' cytotoxic and genotoxic effects were observed only at low concentrations of serum. Copyright © 2011 Wiley Periodicals, Inc.

  10. Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling.

    PubMed

    Shang, Yuqin; Zeng, Yun; Zeng, Yong

    2016-02-02

    Protein glycosylation is one of the key processes that play essential roles in biological functions and dysfunctions. However, progress in glycomics has considerably lagged behind genomics and proteomics, due in part to the enormous challenges in analysis of glycans. Here we present a new integrated and automated microfluidic lectin barcode platform to substantially improve the performance of lectin array for focused glycomic profiling. The chip design and flow control were optimized to promote the lectin-glycan binding kinetics and speed of lectin microarray. Moreover, we established an on-chip lectin assay which employs a very simple blocking method to effectively suppress the undesired background due to lectin binding of antibodies. Using this technology, we demonstrated focused differential profiling of tissue-specific glycosylation changes of a biomarker, CA125 protein purified from ovarian cancer cell line and different tissues from ovarian cancer patients in a fast, reproducible, and high-throughput fashion. Highly sensitive CA125 detection was also demonstrated with a detection limit much lower than the clinical cutoff value for cancer diagnosis. This microfluidic platform holds the potential to integrate with sample preparation functions to construct a fully integrated "sample-to-answer" microsystem for focused differential glycomic analysis. Thus, our technology should present a powerful tool in support of rapid advance in glycobiology and glyco-biomarker development.

  11. Effect of leguminous lectins on the growth of Rhizobium tropici CIAT899.

    PubMed

    de Vasconcelos, Mayron Alves; Cunha, Cláudio Oliveira; Arruda, Francisco Vassiliepe Sousa; Carneiro, Victor Alves; Bastos, Rafaela Mesquita; Mercante, Fábio Martins; do Nascimento, Kyria Santiago; Cavada, Benildo Sousa; dos Santos, Ricardo Pires; Teixeira, Edson Holanda

    2013-05-17

    Rhizobium tropici is a Gram-negative bacterium that induces nodules and fixed atmospheric nitrogen in symbiotic association with Phaseolus vulgaris (common bean) and some other leguminous species. Lectins are proteins that specifically bind to carbohydrates and, consequently, modulate different biological functions. In this study, the d-glucose/ d-mannose-binding lectins (from seeds of Dioclea megacarpa, D. rostrata and D. violacea) and D-galactose-binding lectins (from seeds of Bauhinia variegata, Erythina velutina and Vatairea macrocarpa) were purified using chromatographic techniques and evaluated for their effect on the growth of R. tropici CIAT899. All lectins were assayed with a satisfactory degree of purity according to SDS-PAGE analysis, and stimulated bacterial growth; in particular, the Dioclea rostrata lectin was the most active among all tested proteins. As confirmed in the present study, both d-galactose- and d-glucose/d-mannose-binding lectins purified from the seeds of leguminous plants may be powerful biotechnological tools to stimulate the growth of R. tropici CIAT99, thus improving symbiotic interaction between rhizobia and common bean and, hence, the production of this field crop.

  12. Characterization of storage material in cultured fibroblasts by specific lectin binding in lysosomal storage diseases.

    PubMed

    Virtanen, I; Ekblom, P; Laurila, P; Nordling, S; Raivio, K O; Au