Turboprop Model in the 8- by 6-Foot Supersonic Wind Tunnel

NASA Image and Video Library

1976-08-21

National Aeronautics and Space Administration (NASA) engineer Robert Jeracki prepares a Hamilton Standard SR-1 turboprop model in the test section of the 8- by 6-Foot Supersonic Wind Tunnel at the Lewis Research Center. Lewis researchers were analyzing a series of eight-bladed propellers in their wind tunnels to determine their operating characteristics at speeds up to Mach 0.8. The program, which became the Advanced Turboprop, was part of a NASA-wide Aircraft Energy Efficiency Program which was designed to reduce aircraft fuel costs by 50 percent. The ATP concept was different from the turboprops in use in the 1950s. The modern versions had at least eight blades and were swept back for better performance. After Lewis researchers developed the advanced turboprop theory and established its potential performance capabilities, they commenced an almost decade-long partnership with Hamilton Standard to develop, verify, and improve the concept. A series of 24-inch scale models of the SR-1 with different blade shapes and angles were tested in Lewis’ wind tunnels. A formal program was established in 1978 to examine associated noise levels, aerodynamics, and the drive system. The testing of the large-scale propfan was done on test rigs, in large wind tunnels, and, eventually, on aircraft.

Interior noise control ground test studies for advanced turboprop aircraft applications

NASA Technical Reports Server (NTRS)

Simpson, Myles A.; Cannon, Mark R.; Burge, Paul L.; Boyd, Robert P.

1989-01-01

The measurement and analysis procedures are documented, and the results of interior noise control ground tests conducted on a DC-9 aircraft test section are summarized. The objectives of these tests were to study the fuselage response characteristics of treated and untreated aircraft with aft-mount advanced turboprop engines and to analyze the effectiveness of selected noise control treatments in reducing passenger cabin noise on these aircraft. The results of fuselage structural mode surveys, cabin cavity surveys and sound intensity surveys are presented. The performance of various structural and cabin sidewall treatments is assessed, based on measurements of the resulting interior noise levels under simulated advanced turboprop excitation.

Structural tailoring of advanced turboprops (STAT): User's manual

NASA Technical Reports Server (NTRS)

Brown, K. W.

1991-01-01

This user's manual describes the Structural Tailoring of Advanced Turboprops program. It contains instructions to prepare the input for optimization, blade geometry and analysis, geometry generation, and finite element program control. In addition, a sample input file is provided as well as a section describing special applications (i.e., non-standard input).

Turboprop Cargo Aircraft Systems study, phase 1

NASA Technical Reports Server (NTRS)

Muehlbauer, J. C.; Hewell, J. G., Jr.; Lindenbaum, S. P.; Randall, C. C.; Searle, N.; Stone, F. R., Jr.

1980-01-01

The effects of advanced propellers (propfan) on aircraft direct operating costs, fuel consumption, and noiseprints were determined. A comparison of three aircraft selected from the results with competitive turbofan aircraft shows that advanced turboprop aircraft offer these potential benefits, relative to advanced turbofan aircraft: 21 percent fuel saving, 26 percent higher fuel efficiency, 15 percent lower DOCs, and 25 percent shorter field lengths. Fuel consumption for the turboprop is nearly 40 percent less than for current commercial turbofan aircraft. Aircraft with both types of propulsion satisfy current federal noise regulations. Advanced turboprop aircraft have smaller noiseprints at 90 EPNdB than advanced turbofan aircraft, but large noiseprints at 70 and 80 EPNdB levels, which are usually suggested as quietness goals. Accelerated development of advanced turboprops is strongly recommended to permit early attainment of the potential fuel saving. Several areas of work are identified which may produce quieter turboprop aircraft.

Advanced turboprop aircraft flyover noise: Annoyance to counter-rotating-propeller configurations with an equal number of blades on each rotor, preliminary results

NASA Technical Reports Server (NTRS)

Mccurdy, David A.

1988-01-01

A laboratory experiment was conducted to quantify the annoyance of people to the flyover noise of advanced turboprop aircraft with counter-rotating propellers (CRP) having an equal number of blades on each rotor. The objectives were: to determine the effects of total content on annoyance; and compare annoyance to n x n CRP advanced turboprop aircraft with annoyance to conventional turboprop and jet aircraft. A computer synthesis system was used to generate 27 realistic, time-varying simulations of advanced turboprop takeoff noise in which the tonal content was systematically varied to represent the factorial combinations of nine fundamental frequencies and three tone-to-broadband noise ratios. These advanced turboprop simulations along with recordings of five conventional turboprop takeoffs and five conventional jet takeoffs were presented at three D-weighted sound pressure levels to 64 subjects in an anechoic chamber. Analyses of the subjects' annoyance judgments compare the three aircraft types and examined the effects of the differences in tonal content among the advanced turboprop noises. The annoyance prediction ability of various noise metrics is also examined.

Structural tailoring of advanced turboprops

NASA Technical Reports Server (NTRS)

Brown, K. W.; Hopkins, Dale A.

1988-01-01

The Structural Tailoring of Advanced Turboprops (STAT) computer program was developed to perform numerical optimization on highly swept propfan blades. The optimization procedure seeks to minimize an objective function defined as either: (1) direct operating cost of full scale blade or, (2) aeroelastic differences between a blade and its scaled model, by tuning internal and external geometry variables that must satisfy realistic blade design constraints. The STAT analysis system includes an aerodynamic efficiency evaluation, a finite element stress and vibration analysis, an acoustic analysis, a flutter analysis, and a once-per-revolution forced response life prediction capability. STAT includes all relevant propfan design constraints.

Allison PD 370-42 advanced turboprop engine. Final report, October 1978-February 1979

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stolp, P.

1979-02-01

This study developed data on Detroit Diesel Allison (DDA) common core derivative engines for use in Maritime Patrol Aircraft (MPA) concept formulation studies. The study included the screening of potential DDA turboprop/turboshaft engines and the preparation of technical and planning information on three of the most promising engine candidates plus an all new engine. Screening of DDA Derivative candidates was performed utilizing an analytical MPA model using synthesized mission profiles to rank the candidates in terms of fuel consumption, weight, cost and complexity. The three turboprop engines selected for further study were as follows: a derivative of the unity sizemore » T701-AD-700 shaft power engine with rematched turbine (PD 370-37), an advanced T701 turboprop derivative with 25:1 overall pressure ratio and a scaled ATEGG demonstrated compressor (PD 370-40), an advanced T701 turboprop derivative with 17.7:1 overall pressure ratio and a scaled ATEGG demonstrated compressor (PD 370-4D al and experimental results attests to the accuracy of the assembled mechanism in its description of the homogenrt documents program highlights and research results for CY 1979 along with plans for the completion of program investigations. Postirradiation test data are presented for plateen chemical s.« less

Structural Tailoring of Advanced Turboprops (STAT)

NASA Technical Reports Server (NTRS)

Brown, Kenneth W.

1988-01-01

This interim report describes the progress achieved in the structural Tailoring of Advanced Turboprops (STAT) program which was developed to perform numerical optimizations on highly swept propfan blades. The optimization procedure seeks to minimize an objective function, defined as either direct operating cost or aeroelastic differences between a blade and its scaled model, by tuning internal and external geometry variables that must satisfy realistic blade design constraints. This report provides a detailed description of the input, optimization procedures, approximate analyses and refined analyses, as well as validation test cases for the STAT program. In addition, conclusions and recommendations are summarized.

Study of turboprop systems reliability and maintenance costs

NASA Technical Reports Server (NTRS)

1978-01-01

The overall reliability and maintenance costs (R&MC's) of past and current turboprop systems were examined. Maintenance cost drivers were found to be scheduled overhaul (40%), lack of modularity particularly in the propeller and reduction gearbox, and lack of inherent durability (reliability) of some parts. Comparisons were made between the 501-D13/54H60 turboprop system and the widely used JT8D turbofan. It was found that the total maintenance cost per flight hour of the turboprop was 75% higher than that of the JT8D turbofan. Part of this difference was due to propeller and gearbox costs being higher than those of the fan and reverser, but most of the difference was in the engine core where the older technology turboprop core maintenance costs were nearly 70 percent higher than for the turbofan. The estimated maintenance cost of both the advanced turboprop and advanced turbofan were less than the JT8D. The conclusion was that an advanced turboprop and an advanced turbofan, using similar cores, will have very competitive maintenance costs per flight hour.

Structural Tailoring of Advanced Turboprops (STAT) programmer's manual

NASA Technical Reports Server (NTRS)

Brown, K. W.; Harvey, P. R.

1989-01-01

The Structural Tailoring of Advanced Turboprops (STAT) computer program was developed to perform numerical optimizations on highly swept propfan blades. This manual describes the functionality of the STAT system from a programmer's viewpoint. It provides a top-down description of module intent and interaction. The purpose of this manual is to familiarize the programmer with the STAT system should he/she wish to enhance or verify the program's function.

ATP Interior Noise Technology and Flight Demonstration Program

NASA Technical Reports Server (NTRS)

Stephens, David G.; Powell, Clemans A.

1988-01-01

The paper provides an overview of the ATP (Advanced Turboprop Program) acoustics program with emphasis on the NASA technology program and the recent NASA/Industry demonstration programs aimed at understanding and controlling passenger cabin noise. Technology developments in propeller (source) noise, cabin noise transmission, and subjective acoustics are described. Finally, an overview of the industry demonstrator programs is presented.

Window acoustic study for advanced turboprop aircraft

NASA Technical Reports Server (NTRS)

Prydz, R. A.; Balena, F. J.

1984-01-01

An acoustic analysis was performed to establish window designs for advanced turboprop powered aircraft. The window transmission loss requirements were based on A-weighted interior noise goals of 80 and 75 dBA. The analytical results showed that a triple pane window consisting of two glass outer panes and an inner pane of acrylic would provide the required transmission loss and meet the sidewall space limits. Two window test articles were fabricated for laboratory evaluation and verification of the predicted transmission loss. Procedures for performing laboratory tests are presented.

Advanced turboprop aircraft flyover noise: Annoyance to counter-rotating-propeller configurations with a different number of blades on each rotor: Preliminary results

NASA Technical Reports Server (NTRS)

Mccurdy, David A.

1988-01-01

A laboratory experiment was conducted to quantify the annoyance of people to the flyover noise of advanced turboprop aircraft with counter-rotating propellers (CRP) having a different number of blades on each rotor (nxm, e.g., 10 x 8, 12 x 11). The objectives were: (1) compare annoyance to nxm CRP advanced turboprop aircraft with annoyance to conventional turboprop and jet aircraft; (2) determine the effects of tonal content on annoyance; and (3) determine the ability of aircraft noise measurement procedures and corrections to predict annoyance for this new class of aircraft. A computer synthesis system was used to generate 35 realistic, time-varying simulations of advanced turboprop takeoff noise in which the tonal content was systematically varied to represent combinations of 15 fundamental frequency (blade passage frequency) combinations and three tone-to-broadband noise ratios. The fundamental frequencies, which represented blade number combinations from 6 x 5 to 13 x 12 and 7 x 5 to 13 x 11, ranged from 112.5 to 292.5 Hz. The three tone-to-broadband noise ratios were 0, 15, and 30 dB. These advanced turboprop simulations along with recordings of five conventional turboprop takeoffs and five conventional jet takeoffs were presented at D-weighted sound pressure levels of 70, 80, and 90 dB to 64 subjects in an anechoic chamber. Analyses of the subjects' annoyance judgments compare the three categories of aircraft and examine the effects of the differences in tonal content among the advanced turboprop noises. The annoyance prediction ability of various noise measurement procedures and corrections is also examined.

Comparison of advanced turboprop and turbofan airplanes

NASA Technical Reports Server (NTRS)

Johnson, V. S.

1983-01-01

Results of a parametric study to determine the effects of design variables and penalties on the fuel efficiency of Mach 0.8, 125-passenger, advanced turboprop airplanes show that propeller-wing interference penalty has a major effect. Propeller tip speed has a minor effect, and could be decreased to alleviate the noise problem without significant effects on fuel efficiency. The anticipated noise levels produced by the propfan will require additional acoustical treatment for the fuselage; this additional weight can have a significant effect on fuel efficiency. The propfan advantage over an equivalent technology turbofan is strongly dependent on the interference penalty and acoustical treatment weight. Lowering the cruise Mach number to around 0.73 would result in greatly increased fuel efficiency.

Return of the turboprops

DOE Office of Scientific and Technical Information (OSTI.GOV)

Facey, J.R.; Whitlow, J.B. Jr.; Sievers, G.K.

1988-10-01

Recent advances in propfan development for commercial aircraft cruising at high transonic speeds are discussed in a series of brief overviews and illustrated with photographs and diagrams. The efficiency and performance advantages and the problem of noise are examined; the history of turboprop design since the 1950s is recalled; NASA, DOT, FAA, and DOD turboprop projects are described; and the results of wind-tunnel and flight tests on particular propeller models are reviewed. Current trends in aircraft reconfiguration and acoustic redesign to reduce cabin noise levels are also surveyed.

30 x 60 foot wind tunnel test highlights for an over-the-tail advanced turboprop configuration

NASA Technical Reports Server (NTRS)

Coe, Paul L., Jr.; Perkins, John N.; Rhodes, Graham S.

1991-01-01

This paper presents results from a recent investigation of the static aerodynamic and stability characteristics of a two-surface advanced turboprop aircraft. The conceptual design places Hamilton Standard SR-7 turboprop blades close to the horizontal and vertical tail for potential acoustic shielding. Evaluation of the data shows generally favorable effects of power on aircraft stability and control, and that lateral directional trim can be achieved with one engine inoperative. The tests did show a marked effect of the direction of propeller rotation on thrust minus drag performance.

Comparison ofdvanced turboprop interior noise control ground and flight test data

NASA Technical Reports Server (NTRS)

Simpson, Myles A.; Tran, Boi N.

1992-01-01

Interior noise ground tests conducted on a DC-9 aircraft test section are described. The objectives were to study ground test and analysis techniques for evaluating the effectiveness of interior noise control treatments for advanced turboprop aircraft, and to study the sensitivity of the ground test results to changes in various test conditions. Noise and vibration measurements were conducted under simulated advanced turboprop excitation, for two interior noise control treatment configurations. These ground measurement results were compared with results of earlier UHB (Ultra High Bypass) Demonstrator flight sts with comparable interior treatment configurations. The Demonstrator is an MD-80 test aircraft with the left JT8D engine replaced with a prototype UHB advanced turboprop engine.

Comparison ofdvanced turboprop interior noise control ground and flight test data

NASA Astrophysics Data System (ADS)

Simpson, Myles A.; Tran, Boi N.

Interior noise ground tests conducted on a DC-9 aircraft test section are described. The objectives were to study ground test and analysis techniques for evaluating the effectiveness of interior noise control treatments for advanced turboprop aircraft, and to study the sensitivity of the ground test results to changes in various test conditions. Noise and vibration measurements were conducted under simulated advanced turboprop excitation, for two interior noise control treatment configurations. These ground measurement results were compared with results of earlier UHB (Ultra High Bypass) Demonstrator flight sts with comparable interior treatment configurations. The Demonstrator is an MD-80 test aircraft with the left JT8D engine replaced with a prototype UHB advanced turboprop engine.

Turboprop+: enhanced Turboprop diffusion-weighted imaging with a new phase correction.

PubMed

Lee, Chu-Yu; Li, Zhiqiang; Pipe, James G; Debbins, Josef P

2013-08-01

Faster periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) diffusion-weighted imaging acquisitions, such as Turboprop and X-prop, remain subject to phase errors inherent to a gradient echo readout, which ultimately limits the applied turbo factor (number of gradient echoes between each pair of radiofrequency refocusing pulses) and, thus, scan time reductions. This study introduces a new phase correction to Turboprop, called Turboprop+. This technique employs calibration blades, which generate 2-D phase error maps and are rotated in accordance with the data blades, to correct phase errors arising from off-resonance and system imperfections. The results demonstrate that with a small increase in scan time for collecting calibration blades, Turboprop+ had a superior immunity to the off-resonance-related artifacts when compared to standard Turboprop and recently proposed X-prop with the high turbo factor (turbo factor = 7). Thus, low specific absorption rate and short scan time can be achieved in Turboprop+ using a high turbo factor, whereas off-resonance related artifacts are minimized. © 2012 Wiley Periodicals, Inc.

Allison PD 370-41 derivative turboprop engine. Final report, October 1978-February 1979

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stolp, P.

1979-02-01

This study developed data on Detroit Diesel Allison (DDA) common core derivative engines for use in Maritime Patrol Aircraft (MPA) concept formulation studies. The study included the screening of potential DDA turboprop/turboshaft engines and the preparations of technical and planning information on three of the most promising engine candidates plus an all new engine. Screening of DDA derivative candidates was performed utilizing an analytical MPA model using synthesized mission profiles to rank the candidates in terms of fuel consumption, weight, cost and complexity. The three turboprop engines selected for further study were as follows: a derivative of the unity sizemore » T701-AD-700 shaft power engine with rematched turbine (PD 370-37), and an advanced T701 turboprop derivative with 25:1 overall pressure ratio and a scaled ATEGG demonstrated compressor (PD 370-40), an advanced T701 turboprop derivative with 17.7:1 overall pressure ratio and a scaled ATEGG demonstrated compressor.« less

Study of cost/benefit tradeoffs for reducing the energy consumption of the commercial air transportation system. [Advanced turboprop introduction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Coykendall, R.E.; Curry, J.K.; Domke, A.E.

1976-06-01

Economic studies were conducted for three general fuel-conserving options: (1) improving fuel-consumption characteristics of existing aircraft via retrofit modifications; (2) introducing fuel-efficient derivations of existing production aircraft and/or introducing fuel efficient, current state-of-the-art new aircraft; and (3) introducing an advanced state-of-the-art turboprop airplane. These studies were designed to produce an optimum airline fleet mix for the years 1980, 1985 and 1990. The fleet selected accommodated a normal growth market by introducing somewhat larger aircraft while solving for maximum departure frequencies and a minimum load factor corresponding to a 15% investment hurdle rate. Fuel burnt per available-seat-mile flown would drop 22%more » from 1980 to 1990 due to the use of more fuel efficient aircraft designs, larger average aircraft size, and increased seating density. An inflight survey was taken to determine air traveler attitudes towards a new generation of advanced turboprops. (Author) (GRA)« less

Structural Tailoring of Advanced Turboprops (STAT). Theoretical manual

NASA Technical Reports Server (NTRS)

Brown, K. W.

1992-01-01

This manual describes the theories in the Structural Tailoring of Advanced Turboprops (STAT) computer program, which was developed to perform numerical optimizations on highly swept propfan blades. The optimization procedure seeks to minimize an objective function, defined as either direct operating cost or aeroelastic differences between a blade and its scaled model, by tuning internal and external geometry variables that must satisfy realistic blade design constraints. The STAT analyses include an aerodynamic efficiency evaluation, a finite element stress and vibration analysis, an acoustic analysis, a flutter analysis, and a once-per-revolution (1-p) forced response life prediction capability. The STAT constraints include blade stresses, blade resonances, flutter, tip displacements, and a 1-P forced response life fraction. The STAT variables include all blade internal and external geometry parameters needed to define a composite material blade. The STAT objective function is dependent upon a blade baseline definition which the user supplies to describe a current blade design for cost optimization or for the tailoring of an aeroelastic scale model.

Structural Tailoring of Advanced Turboprops (STAT). Theoretical manual

NASA Astrophysics Data System (ADS)

Brown, K. W.

1992-10-01

This manual describes the theories in the Structural Tailoring of Advanced Turboprops (STAT) computer program, which was developed to perform numerical optimizations on highly swept propfan blades. The optimization procedure seeks to minimize an objective function, defined as either direct operating cost or aeroelastic differences between a blade and its scaled model, by tuning internal and external geometry variables that must satisfy realistic blade design constraints. The STAT analyses include an aerodynamic efficiency evaluation, a finite element stress and vibration analysis, an acoustic analysis, a flutter analysis, and a once-per-revolution (1-p) forced response life prediction capability. The STAT constraints include blade stresses, blade resonances, flutter, tip displacements, and a 1-P forced response life fraction. The STAT variables include all blade internal and external geometry parameters needed to define a composite material blade. The STAT objective function is dependent upon a blade baseline definition which the user supplies to describe a current blade design for cost optimization or for the tailoring of an aeroelastic scale model.

The Langley turbo-prop commuter design: A complete project description

NASA Technical Reports Server (NTRS)

Buttram, Greg; Horton, Keith; Keeter, Tim; Millhouse, Paul; Newberry, Kelli; Obyrne, Brian

1991-01-01

The primary objective of this project was to propose and prove the possibility of a new, advanced technology commuter aircraft design. Among the specifications were short to medium range capabilities, low seat per mile cost, fuel efficiency, and passenger comfort. Based on market evaluation, we found that the optimum size for new regional aircraft is around 50 passengers; we have designed our aircraft for this capacity. Turboprop engines provide substantial reductions in operating costs due to lower fuel consumption. We have therefore chosen an advanced turboprop engine. Composite materials, while more expensive to purchase and manufacture, result in decreased costs later through weight savings and ease of replacement.

Preliminary study of advanced turboprop and turboshaft engines for light aircraft. [cost effectiveness

NASA Technical Reports Server (NTRS)

Knip, G.; Plencner, R. M.; Eisenberg, J. D.

1980-01-01

The effects of engine configuration, advanced component technology, compressor pressure ratio and turbine rotor-inlet temperature on such figures of merit as vehicle gross weight, mission fuel, aircraft acquisition cost, operating, cost and life cycle cost are determined for three fixed- and two rotary-wing aircraft. Compared with a current production turboprop, an advanced technology (1988) engine results in a 23 percent decrease in specific fuel consumption. Depending on the figure of merit and the mission, turbine engine cost reductions required to achieve aircraft cost parity with a current spark ignition reciprocating (SIR) engine vary from 0 to 60 percent and from 6 to 74 percent with a hypothetical advanced SIR engine. Compared with a hypothetical turboshaft using currently available technology (1978), an advanced technology (1988) engine installed in a light twin-engine helicopter results in a 16 percent reduction in mission fuel and about 11 percent in most of the other figures of merit.

An estimate of the enroute noise of an advanced turboprop airplane NASA-TM-87302 E-3020 NAS 1.15:87302 HC A02/MF A01

NASA Technical Reports Server (NTRS)

Dittmar, J. H.

1986-01-01

The enroute noise of an Advanced Turboprop powered aircraft was estimated. The measured noise levels were roughly equivalent in annoyance to the noise 15.24 m from an automobile traveling at 80 km/h. It is felt that these levels would not illicit noise complaints from urban areas during the day but might be a slight annoyance in rural areas or in urban areas at night. Although it is not felt that the enroute noise is a major problem, it is indicated that a reduction in the enroute noise could improve the acceptability of advance turboprop airplanes.

Acoustic test and analyses of three advanced turboprop models

NASA Technical Reports Server (NTRS)

Brooks, B. M.; Metzger, F. B.

1980-01-01

Results of acoustic tests of three 62.2 cm (24.5 inch) diameter models of the prop-fan (a small diameter, highly loaded. Multi-bladed variable pitch advanced turboprop) are presented. Results show that there is little difference in the noise produced by unswept and slightly swept designs. However, the model designed for noise reduction produces substantially less noise at test conditions simulating 0.8 Mach number cruise speed or at conditions simulating takeoff and landing. In the near field at cruise conditions the acoustically designed. In the far field at takeoff and landing conditions the acoustically designed model is 5 db quieter than unswept or slightly swept designs. Correlation between noise measurement and theoretical predictions as well as comparisons between measured and predicted acoustic pressure pulses generated by the prop-fan blades are discussed. The general characteristics of the pulses are predicted. Shadowgraph measurements were obtained which showed the location of bow and trailing waves.

Wright Field turboprop study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lorenzetti, R.C.; Dull, P.P.

1981-01-01

The considered investigation was conducted as part of an effort to reduce aircraft fuel costs. Turboprops are very fuel efficient at speeds below Mach 0.6. One of the most promising approaches to reduce fuel consumption in the speed range from Mach 0.6 to 0.8 is related to an employment of the swept eight to ten blade propeller (propfan) being developed by NASA. All studies to date indicate that turboshaft engines with propfans might save 15% or more of the fuel used by a turbofan engine of comparable technology. Attention is given to a turboprop status review, contractor studies, the NASAmore » propfan program, an in-house performance study, specific problems which have to be solved to make a twin-turboprop, 100-150 passenger transport a viable possibility for the early 1990s, and turboprop dilemmas.« less

Turboprop aircraft against terrorism: a SWOT analysis of turboprop aircraft in CAS operations

NASA Astrophysics Data System (ADS)

Yavuz, Murat; Akkas, Ali; Aslan, Yavuz

2012-06-01

Today, the threat perception is changing. Not only for countries but also for defence organisations like NATO, new threat perception is pointing terrorism. Many countries' air forces become responsible of fighting against terorism or Counter-Insurgency (COIN) Operations. Different from conventional warfare, alternative weapon or weapon systems are required for such operatioins. In counter-terrorism operations modern fighter jets are used as well as helicopters, subsonic jets, Unmanned Aircraft Systems (UAS), turboprop aircraft, baloons and similar platforms. Succes and efficiency of the use of these platforms can be determined by evaluating the conditions, the threats and the area together. Obviously, each platform has advantages and disadvantages for different cases. In this research, examples of turboprop aircraft usage against terrorism and with a more general approach, turboprop aircraft for Close Air Support (CAS) missions from all around the world are reviewed. In this effort, a closer look is taken at the countries using turboprop aircraft in CAS missions while observing the fields these aircraft are used in, type of operations, specifications of the aircraft, cost and the maintenance factors. Thus, an idea about the convenience of using these aircraft in such operations can be obtained. A SWOT analysis of turboprop aircraft in CAS operations is performed. This study shows that turboprop aircraft are suitable to be used in counter-terrorism and COIN operations in low threat environment and is cost benefical compared to jets.

Potential reduction of en route noise from an advanced turboprop aircraft

NASA Technical Reports Server (NTRS)

Dittmar, James H.

1990-01-01

When the en route noise of a representative aircraft powered by an eight-blade SR-7 propeller was previously calculated, the noise level was cited as a possible concern associated with the acceptance of advanced turboprop aircraft. Some potential methods for reducing the en route noise were then investigated and are reported. Source noise reductions from increasing the blade number and from operating at higher rotative speed to reach a local minimum noise point were investigated. Greater atmospheric attenuations for higher blade passing frequencies were also indicated. Potential en route noise reductions from these methods were calculated as 9.5 dB (6.5 dB(A)) for a 10-blade redesigned propeller and 15.5 dB (11 dB(A)) for a 12-blade redesigned propeller.

Exploratory wind tunnel investigation of the stability and control characteristics of a three-surface, forward-swept wing advanced turboprop model

NASA Technical Reports Server (NTRS)

Coe, Paul L., Jr.; Perkins, John N.; Owens, D. Bruce

1990-01-01

The purpose of the present investigation was to parametrically study the stability and control characteristics of a forward-swept wing three-surface turboprop model through an extended angle of attack range, including the deep-stall region. As part of a joint research program between North Carolina State University and NASA Langley Research Center, a low-speed wind tunnel investigation was conducted with a three-surface, forward-swept wing, aft-mounted, twin-pusher propeller, model, representative of an advanced turboprop configuration. The tests were conducted in the NASA Langley 12-Foot Low-Speed Wind Tunnel. The model parameters varied in the test were horizontal tail location, canard size, sweep and location, and wing position. The model was equipped with air turbines, housed within the nacelles and driven by compressed air, to model turboprop power effects. A three-surface, forward-swept wing configuration that provided satisfactory static longitudinal and lateral/directional stability was identified. The three-surface configuration was found to have greater longitudinal control and increased center of gravity range relative to a conventional (two-surface) design. The test showed that power had a large favorable effect on stability and control about all three axis in the post-stall regime.

Advanced turbo-prop airplane interior noise reduction-source definition

NASA Technical Reports Server (NTRS)

Magliozzi, B.; Brooks, B. M.

1979-01-01

Acoustic pressure amplitudes and phases were measured in model scale on the surface of a rigid semicylinder mounted in an acoustically treated wind tunnel near a prop-fan (an advanced turboprop with many swept blades) model. Operating conditions during the test simulated those of a prop-fan at 0.8 Mach number cruise. Acoustic pressure amplitude and phase contours were defined on the semicylinder surface. Measurements obtained without the semi-cylinder in place were used to establish the magnitude of pressure doubling for an aircraft fuselage located near a prop-fan. Pressure doubling effects were found to be 6dB at 90 deg incidence decreasing to no effect at grazing incidence. Comparisons of measurements with predictions made using a recently developed prop-fan noise prediction theory which includes linear and non-linear source terms showed good agreement in phase and in peak noise amplitude. Predictions of noise amplitude and phase contours, including pressure doubling effects derived from test, are included for a full scale prop-fan installation.

Fundamental studies of structure borne noise for advanced turboprop applications

NASA Technical Reports Server (NTRS)

Eversman, W.; Koval, L. R.

1985-01-01

The transmission of sound generated by wing-mounted, advanced turboprop engines into the cabin interior via structural paths is considered. The structural model employed is a beam representation of the wing box carried into the fuselage via a representative frame type of carry through structure. The structure for the cabin cavity is a stiffened shell of rectangular or cylindrical geometry. The structure is modelled using a finite element formulation and the acoustic cavity is modelled using an analytical representation appropriate for the geometry. The structural and acoustic models are coupled by the use of hard wall cavity modes for the interior and vacuum structural modes for the shell. The coupling is accomplished using a combination of analytical and finite element models. The advantage is the substantial reduction in dimensionality achieved by modelling the interior analytically. The mathematical model for the interior noise problem is demonstrated with a simple plate/cavity system which has all of the features of the fuselage interior noise problem.

Predicted changes in advanced turboprop noise with shaft angle of attack

NASA Technical Reports Server (NTRS)

Padula, S. L.; Block, P. J. W.

1984-01-01

Advanced turboprop blade designs and new propeller installation schemes motivated an effort to include unsteady loading effects in existing propeller noise prediction computer programs. The present work validates the prediction capability while studing the effects of shaft inclination on the radiated sound field. Classical methods of propeller performance analysis supply the time-dependent blade loading needed to calculate noise. Polar plots of the sound pressure level (SPL) of the first four harmonics and overall SPL are indicative of the change in directivity pattern as a function of propeller angle of attack. Noise predictions are compared with newly available wind tunnel data and the accuracy and applicability of the prediction method are discussed. It is concluded that unsteady blade loading caused by inclining the propeller with respect to the flow changes the directionality and the intensity of the radiated noise. These changes are well modeled by the present quasi-steady prediction method.

Laboratory experiments on active suppression of advanced turboprop noise

NASA Technical Reports Server (NTRS)

Dittmar, J. H.

1985-01-01

The noise generated by supersonic tip speed propellers may be a cabin environment problem for future propeller-driven airplanes. Active suppression from speakers inside the airplane cabin has been proposed for canceling out this noise. The potential of active suppression of advanced turboprop noise was tested by using speakers in a rectangular duct. Experiments were first performed with sine wave signals. The results compared well with the ideal cancellation curve of noise as a function of phase angle. Recorded noise signals from subsonic and supersonic tip speed propellers were than used in the duct to deterthe potential for canceling their noise. The subsonic propeller data showed significant cancellations but less than those obtained with the sine wave. The blade-passing-tone cancellation curve for the supersonic propeller was very similar to the subsonic curve, indicating that it is potentially just as easy to cancel supersonic as subsonic propeller blade-passing-tone noise. Propeller duct data from a recorded propeller source and spatial data taken on a propeller-drive airplane showed generally good agreement when compared versus phase angle. This agreement, combined with the similarity of the subsonic and supersonic duct propeller data, indicates that the area of cancellation for advanced supersonic propellers will be similar to that measured on the airplane. Since the area of cancellation on the airplane was small, a method for improving the active noise suppression by using outside speakers is discussed.

Results of the pollution reduction technology program for turboprop engines

NASA Technical Reports Server (NTRS)

Mularz, E. J.

1976-01-01

A program was performed to evolve and demonstrate advanced combustor technology aimed at achieving the 1979 EPA standards for turboprop engines (Class P2). The engine selected for this program was the 501-D22A turboprop. Three combustor concepts were designed and tested in a combustor rig at the exact combustor operating conditions of the 50-D22A engine over the EPA landing-takeoff cycle. Each combustor concept exhibited pollutant emissions well below the EPA standards, achieving substantial reductions in unburned hydrocarbons, carbon monoxide, and smoke emissions compared with emissions from the production combustor of this engine. Oxides of nitrogen emissions remained well below the EPA standards, also.

Low-speed wind-tunnel investigation of the flight dynamic characteristics of an advanced turboprop business/commuter aircraft configuration

NASA Technical Reports Server (NTRS)

Coe, Paul L., Jr.; Turner, Steven G.; Owens, D. Bruce

1990-01-01

An investigation was conducted to determine the low-speed flight dynamic behavior of a representative advanced turboprop business/commuter aircraft concept. Free-flight tests were conducted in the NASA Langley Research Center's 30- by 60-Foot Tunnel. In support of the free-flight tests, conventional static, dynamic, and free-to-roll oscillation tests were performed. Tests were intended to explore normal operating and post stall flight conditions, and conditions simulating the loss of power in one engine.

Analysis of results from wind tunnel tests of inlets for an advanced turboprop nacelle installation

NASA Technical Reports Server (NTRS)

Hancock, J. P.; Lyman, V.; Pennock, A. P.

1986-01-01

Inlets for tractor installations of advanced turboprop propulsion systems were tested in three phases, covering a period from November, 1982 to January, 1984. Nacelle inlet configuration types included single scoop, twin scoop, and annular arrangements. Tests were performed with and without boundary layer diverters and several different diverter heights were tested for the single scoop inlet. This same inlet was also tested at two different axial positions. Test Mach numbers ranged from Mach 0.20 to 0.80. Types of data taken were: (1) internal and external pressures, including inlet throat recoveries; (2) balance forces, including thrust-minus-drag; and (3) propellar blade stresses.

Pollution Reduction Technology Program, Turboprop Engines, Phase 1

NASA Technical Reports Server (NTRS)

Anderson, R. D.; Herman, A. S.; Tomlinson, J. G.; Vaught, J. M.; Verdouw, A. J.

1976-01-01

Exhaust pollutant emissions were measured from a 501-D22A turboprop engine combustor and three low emission combustor types -- reverse flow, prechamber, and staged fuel, operating over a fuel-air ratio range of .0096 to .020. The EPAP LTO cycle data were obtained for a total of nineteen configurations. Hydrocarbon emissions were reduced from 15.0 to .3 lb/1000 Hp-Hr/cycle, CO from 31.5 to 4.6 lb/1000 Hp-Hr/cycle with an increase in NOx of 17 percent, which is still 25% below the program goal. The smoke number was reduced from 59 to 17. Emissions given here are for the reverse flow Mod. IV combustor which is the best candidate for further development into eventual use with the 501-D22A turboprop engine. Even lower emissions were obtained with the advanced technology combustors.

Measured far-field flight noise of a counterrotation turboprop at cruise conditions

NASA Technical Reports Server (NTRS)

Woodward, Richard P.; Loeffler, Irvin J.; Dittmar, James H.

1989-01-01

Modern high speed propeller (advanced turboprop) aircraft are expected to operate on 50 to 60 percent less fuel than the 1980 vintage turbofan fleet while at the same time matching the flight speed and performance of those aircraft. Counterrotation turboprop engines offer additional fuel savings by means of upstream propeller swirl recovery. This paper presents acoustic sideline results for a full-scale counterrotation turboprop engine at cruise conditions. The engine was installed on a Boeing 727 aircraft in place of the right-side turbofan engine. Acoustic data were taken from an instrumented Learjet chase plane. Sideline acoustic results are presented for 0.50 and 0.72 Mach cruise conditions. A scale model of the engine propeller was tested in a wind tunnel at 0.72 Mach cruise conditions. The model data were adjusted to flight acquisition conditions and were in general agreement with the flight results.

User's guide to PMESH: A grid-generation program for single-rotation and counterrotation advanced turboprops

NASA Technical Reports Server (NTRS)

Warsi, Saif A.

1989-01-01

A detailed operating manual is presented for a grid generating program that produces 3-D meshes for advanced turboprops. The code uses both algebraic and elliptic partial differential equation methods to generate single rotation and counterrotation, H or C type meshes for the z - r planes and H type for the z - theta planes. The code allows easy specification of geometrical constraints (such as blade angle, location of bounding surfaces, etc.), mesh control parameters (point distribution near blades and nacelle, number of grid points desired, etc.), and it has good runtime diagnostics. An overview is provided of the mesh generation procedure, sample input dataset with detailed explanation of all input, and example meshes.

A new generation T56 turboprop engine

DOE Office of Scientific and Technical Information (OSTI.GOV)

McIntire, W.L.

1984-06-01

The focus of the T56 Series IV turboprop engine development program is to improve power and fuel consumption through incorporation of demonstrated technology improvements while retaining the long term durability and cost effective design of the T56 family. The T56-A-427, the Navy Series IV derivative of the 5000 shp (3728.5 kW) class T56 turboprop engine, resulted from over ten years of technology development via Advanced Turbine Engine Gas Generator (ATEGG), Joint Technology Demonstrator Engine (JTDE), and advanced component programs at Allison Gas Turbine Operations. An example of government and industry cooperation to transfer advanced gas turbine technology is the Airmore » Force Engine Model Derivative Program (EMDP). The initial full-scale demonstration in this program confirmed a 10-1/2% reduction in specific fuel consumption (sfc) and a power growth of 21% in the basic T56 frame. Continued early demonstrations and development by IR and D, Navy funds, and Allison discretionary funds showed a further sfc reduction to 13% and power increase of 28%. The full-scale development program is now underway to provide production engines in late 1986. Engines will be available for the Grumman E-2 and C-2 aircraft, with follow-on adaptions for Lockheed C-130/L100 and P-3 aircraft, and generator sets for DD 963, DDG 993, CG 47 and DDG 51 warships.« less

Turboprop cargo aircraft systems study

NASA Technical Reports Server (NTRS)

Muehlbauer, J. C.; Hewell, J. G., Jr.; Lindenbaum, S. P.; Randall, C. C.; Searle, N.; Stone, R. G., Jr.

1981-01-01

The effects of using advanced turboprop propulsion systems to reduce the fuel consumption and direct operating costs of cargo aircraft were studied, and the impact of these systems on aircraft noise and noise prints around a terminal area was determined. Parametric variations of aircraft and propeller characteristics were investigated to determine their effects on noiseprint areas, fuel consumption, and direct operating costs. From these results, three aircraft designs were selected and subjected to design refinements and sensitivity analyses. Three competitive turbofan aircraft were also defined from parametric studies to provide a basis for comparing the two types of propulsion.

Pollution reduction technology program for turboprop engines

NASA Technical Reports Server (NTRS)

Tomlinson, J. G.

1977-01-01

The reduction of CO, HC, and smoke emissions while maintaining acceptable NO(x) emissions without affecting fuel consumption, durability, maintainability, and safety was accomplished. Component combustor concept screening directed toward the demonstration of advanced combustor technology required to meet the EPA exhaust emissions standards for class P2 turboprop engines was covered. The combustion system for the Allison 501-D22A engine was used, and three combustor design concepts - reverse flow, prechamber, and staged fuel were evaluated.

Motion Correction in PROPELLER and Turboprop-MRI

PubMed Central

Tamhane, Ashish A.; Arfanakis, Konstantinos

2009-01-01

PROPELLER and Turboprop-MRI are characterized by greatly reduced sensitivity to motion, compared to their predecessors, fast spin-echo and gradient and spin-echo, respectively. This is due to the inherent self-navigation and motion correction of PROPELLER-based techniques. However, it is unknown how various acquisition parameters that determine k-space sampling affect the accuracy of motion correction in PROPELLER and Turboprop-MRI. The goal of this work was to evaluate the accuracy of motion correction in both techniques, to identify an optimal rotation correction approach, and determine acquisition strategies for optimal motion correction. It was demonstrated that, blades with multiple lines allow more accurate estimation of motion than blades with fewer lines. Also, it was shown that Turboprop-MRI is less sensitive to motion than PROPELLER. Furthermore, it was demonstrated that the number of blades does not significantly affect motion correction. Finally, clinically appropriate acquisition strategies that optimize motion correction were discussed for PROPELLER and Turboprop-MRI. PMID:19365858

Interior noise considerations for advanced high-speed turboprop aircraft

NASA Technical Reports Server (NTRS)

Mixson, J. S.; Farassat, F.; Leatherwood, J. D.; Prydz, R.; Revell, J. D.

1982-01-01

This paper describes recent research on noise generated by high-speed propellers, on noise transmission through acoustically treated aircraft sidewalls and on subjective response to simulated turboprop noise. Propeller noise discussion focuses on theoretical prediction methods for complex blade shapes designed for low noise at Mach = 0.8 flight and on comparisons with experimental test results. Noise transmission experiments using a 168 cm. diameter aircraft fuselage model and scaled heavy-double-wall treatments indicate that the treatments perform well and that the predictions are usually conservative. Studies of subjective comfort response in an anechoic environment are described for noise signatures having combinations of broadband and propeller-type tone components.

Small Engine Technology (SET) - Task 4, Regional Turboprop/Turbofan Engine Advanced Combustor Study

NASA Technical Reports Server (NTRS)

Reynolds, Robert; Srinivasan, Ram; Myers, Geoffrey; Cardenas, Manuel; Penko, Paul F. (Technical Monitor)

2003-01-01

Under the SET Program Task 4 - Regional Turboprop/Turbofan Engine Advanced Combustor Study, a total of ten low-emissions combustion system concepts were evaluated analytically for three different gas turbine engine geometries and three different levels of oxides of nitrogen (NOx) reduction technology, using an existing AlliedSignal three-dimensional (3-D) Computational Fluid Dynamics (CFD) code to predict Landing and Takeoff (LTO) engine cycle emission values. A list of potential Barrier Technologies to the successful implementation of these low-NOx combustor designs was created and assessed. A trade study was performed that ranked each of the ten study configurations on the basis of a number of manufacturing and durability factors, in addition to emissions levels. The results of the trade study identified three basic NOx-emissions reduction concepts that could be incorporated in proposed follow-on combustor technology development programs aimed at demonstrating low-NOx combustor hardware. These concepts are: high-flow swirlers and primary orifices, fuel-preparation cans, and double-dome swirlers.

Application of advanced high speed turboprop technology to future civil short-haul transport aircraft design

NASA Technical Reports Server (NTRS)

Conlon, J. A.; Bowles, J. V.

1978-01-01

With an overall goal of defining the needs and requirements for short-haul transport aircraft research and development, the objective of this paper is to determine the performance and noise impact of short-haul transport aircraft designed with an advanced turboprop propulsion system. This propulsion system features high-speed propellers that have more blades and reduced diameters. Aircraft are designed for short and medium field lengths; mission block fuel and direct operating costs (DOC) are used as performance measures. The propeller diameter was optimized to minimize DOC. Two methods are employed to estimate the weight of the acoustic treatment needed to reduce interior noise to an acceptable level. Results show decreasing gross weight, block fuel, DOC, engine size, and optimum propfan diameter with increasing field length. The choice of acoustic treatment method has a significant effect on the aircraft design.

Evaluation of a staged fuel combustor for turboprop engines

NASA Technical Reports Server (NTRS)

Verdouw, A. J.

1976-01-01

Proposed EPA emission regulations require emission reduction by 1979 for various gas turbine engine classes. Extensive combustion technology advancements are required to meet the proposed regulations. The T56 turboprop engine requires CO, UHC, and smoke reduction. A staged fuel combustor design was tested on a combustion rig to evaluate emission reduction potential in turboprop engines from fuel zoning. The can-type combustor has separately fueled-pilot and main combustion zones in series. The main zone fueling system was arranged for potential incorporation into the T56 with minor or no modifications to the basic engine. Three combustor variable geometry systems were incorporated to evaluate various airflow distributions. Emission results with fixed geometry operation met all proposed EPA regulations over the EPA LTO cycle. CO reduction was 82 percent, UHC reduction was 96 percent, and smoke reduction was 84 percent. NOx increased 14 percent over the LTO cycle. At high power, NOx reduction was 40 to 55 percent. This NOx reduction has potential application to stationary gas turbine powerplants which have different EPA regulations.

ANP TURBOPROP STUDIES BIBLIOGRAPHY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cope, A.D. comp.

1957-04-01

Documents, drawings, photographs, slides, and charts issued by ANPD and reports and drawings by AGT-FPLD, Lockheed, and Douglas Aircraft Company on ANP turboprop studies from April 8, 1955 to March 31, 1957 are listed. (auth)

Turboprop Propulsion Mechanic.

ERIC Educational Resources Information Center

Chanute AFB Technical Training Center, IL.

This instructional package consists of a plan of instruction, glossary, and student handouts and exercises for use in training Air Force personnel to become turboprop propulsion mechanics. Addressed in the individual lessons of the course are the following: common hand tools, hardware, measuring devices, and safety wiring; aircraft and engine…

Analysis and test evaluation of the dynamic response and stability of three advanced turboprop models at low forward speed

NASA Technical Reports Server (NTRS)

Smith, Arthur F.

1985-01-01

Results of wind tunnel tests at low forward speed for blade dynamic response and stability of three 62.2 cm (24.5 in) diameter models of the Prop-Fan, advanced turboprop, are presented. Measurements of dynamic response were made with the rotors mounted on an isolated nacelle, with varying tilt for nonuniform inflow. Low speed stall flutter tests were conducted at Mach numbers from 0.0 to 0.35. Measurements are compared to Eigen-solution flutter boundaries. Calculated 1P stress response agrees favorably with experiment. Predicted stall flutter boundaries correlate well with measured high stress regions. Stall flutter is significantly reduced by increased blade sweep. Susceptibility to stall flutter decreases rapidly with forward speed.

Low-speed stability and control characteristics of a transport model with aft-fuselage-mounted advanced turboprops

NASA Technical Reports Server (NTRS)

Applin, Z. T.; Coe, P. L., Jr.

1986-01-01

A limited experimental investigation was conducted in the Langley 4- by 7-Meter Tunnel to explore the effects of aft-fuselage-mounted advanced turboprop installations on the low-speed stability and control characteristics of a representative transport aircraft in a landing configuration. In general, the experimental results indicate that the longitudinal and lateral-directional stability characteristics for the aft-fuselage-mounted single-rotation tractor and counter-rotation pusher propeller configurations tested during this investigation are acceptable aerodynamically. For the single-rotation tractor configuration, the propeller-induced aerodynamics are significantly influenced by the interaction of the propeller slipstream with the pylon and nacelle. The stability characteristics for the counter-rotation pusher configuration are strongly influenced by propeller normal forces. The longitudinal and directional control effectiveness, engine-out characteristics, and ground effects are also presented. In addition, a tabulated presentation of all aerodynamic data presented in this report is included as an appendix.

77 FR 56794 - Airworthiness Directives; Pratt & Whitney Canada Corp. Turboprop Engines

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-14

... Canada Corp. Turboprop Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of..., PW127F, PW127G, and PW127M turboprop engines. The existing AD currently requires initial and repetitive... 56795

76 FR 61255 - Airworthiness Directives; Honeywell International Inc. TPE331 Model Turboprop Engines With...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-04

... Airworthiness Directives; Honeywell International Inc. TPE331 Model Turboprop Engines With Certain Dixie... Honeywell International Inc. TPE331 model turboprop engines with a part manufacturer approval (PMA... all Honeywell International Inc. TPE331 model turboprop engines with a PMA replacement Dixie Aerospace...

En route noise of two turboprop aircraft

NASA Technical Reports Server (NTRS)

Dobrzynski, Werner

1990-01-01

In order to weigh en route noise emissions originating from future propfan powered aircraft, a data base of emission levels from conventional turboprop aircraft is needed. For this reason flyover noise measurements on two twin-engine turboprop aircraft were conducted at flight heights between 17,000 and 21,000 ft. Acoustic data are presented together with propeller operational parameters and environmental meteorological data. Narrowband spectral analyses demonstrate the characteristic features of the measured propeller noise signatures: Noise spectra are dominated by the propeller rotational noise fundamental frequency and pronounced noise beats occur as a consequence of different rotational speeds of the propellers.

Analysis and test evaluation of the dynamic response and stability of three advanced turboprop models

NASA Technical Reports Server (NTRS)

Bansal, P. N.; Arseneaux, P. J.; Smith, A. F.; Turnberg, J. E.; Brooks, B. M.

1985-01-01

Results of dynamic response and stability wind tunnel tests of three 62.2 cm (24.5 in) diameter models of the Prop-Fan, advanced turboprop, are presented. Measurements of dynamic response were made with the rotors mounted on an isolated nacelle, with varying tilt for nonuniform inflow. One model was also tested using a semi-span wing and fuselage configuration for response to realistic aircraft inflow. Stability tests were performed using tunnel turbulence or a nitrogen jet for excitation. Measurements are compared with predictions made using beam analysis methods for the model with straight blades, and finite element analysis methods for the models with swept blades. Correlations between measured and predicted rotating blade natural frequencies for all the models are very good. The IP dynamic response of the straight blade model is reasonably well predicted. The IP response of the swept blades is underpredicted and the wing induced response of the straight blade is overpredicted. Two models did not flutter, as predicted. One swept blade model encountered an instability at a higher RPM than predicted, showing predictions to be conservative.

Real-time Simulation of Turboprop Engine Control System

NASA Astrophysics Data System (ADS)

Sheng, Hanlin; Zhang, Tianhong; Zhang, Yi

2017-05-01

On account of the complexity of turboprop engine control system, real-time simulation is the technology, under the prerequisite of maintaining real-time, to effectively reduce development cost, shorten development cycle and avert testing risks. The paper takes RT-LAB as a platform and studies the real-time digital simulation of turboprop engine control system. The architecture, work principles and external interfaces of RT-LAB real-time simulation platform are introduced firstly. Then based on a turboprop engine model, the control laws of propeller control loop and fuel control loop are studied. From that and on the basis of Matlab/Simulink, an integrated controller is designed which can realize the entire process control of the engine from start-up to maximum power till stop. At the end, on the basis of RT-LAB platform, the real-time digital simulation of the designed control system is studied, different regulating plans are tried and more ideal control effects have been obtained.

A hybrid numerical technique for predicting the aerodynamic and acoustic fields of advanced turboprops

NASA Technical Reports Server (NTRS)

Homicz, G. F.; Moselle, J. R.

1985-01-01

A hybrid numerical procedure is presented for the prediction of the aerodynamic and acoustic performance of advanced turboprops. A hybrid scheme is proposed which in principle leads to a consistent simultaneous prediction of both fields. In the inner flow a finite difference method, the Approximate-Factorization Alternating-Direction-Implicit (ADI) scheme, is used to solve the nonlinear Euler equations. In the outer flow the linearized acoustic equations are solved via a Boundary-Integral Equation (BIE) method. The two solutions are iteratively matched across a fictitious interface in the flow so as to maintain continuity. At convergence the resulting aerodynamic load prediction will automatically satisfy the appropriate free-field boundary conditions at the edge of the finite difference grid, while the acoustic predictions will reflect the back-reaction of the radiated field on the magnitude of the loading source terms, as well as refractive effects in the inner flow. The equations and logic needed to match the two solutions are developed and the computer program implementing the procedure is described. Unfortunately, no converged solutions were obtained, due to unexpectedly large running times. The reasons for this are discussed and several means to alleviate the situation are suggested.

Design and analysis of a fuel-efficient single-engine, turboprop-powered, business airplane

NASA Technical Reports Server (NTRS)

Martin, G. L.; Everest, D. E., Jr.; Lovell, W. A.; Price, J. E.; Walkley, K. B.; Washburn, G. F.

1981-01-01

The speed, range, payload, and fuel efficiency of a general aviation airplane powered by one turboprop engine was determined and compared to a twin engine turboprop aircraft. An airplane configuration was developed which can carry six people for a noreserve range of 2,408 km at a cruise speed above 154 m/s, and a cruise altitude of about 9,144 m. The cruise speed is comparable to that of the fastest of the current twin turboprop powered airplanes. It is found that the airplane has a cruise specific range greater than all twin turboprop engine airplanes flying in its speed range and most twin piston engine airplanes flying at considerably slower cruise airspeeds.

The Bimotor Turboprop Multi-Purpose Aircraft, the Yun-11T (Y-11),

DTIC Science & Technology

1982-08-13

TRANSLATION FTD-ID(RS)T-0521-82 13 August 1982 MICROFICHE N : FrD-82-C-001105 THE BIMOTOR TURBOPROP MULTI-PURPOSE AIRCRAFT, THE YUll 11T ( Y -11) By...AD-A142 848 THE BIMOTOR TURBOPROP MULTI-PURPOSE AIRCRAFT THE i/i YUN-jiT ( Y -ii)(U) FOREIGN TECHNOLOGY DIV WRIGHT-PATTERSON RFB OH X WENJIE 03 AUG 82...BUREAU OF STANDARDS- 1963-A -. -q 4 ,,> 1 .77. ADA1 42848 -. -,. FOREIGN TECHNOLOGY DIVISION THE BIMOTOR TURBOPROP MULTI-PURPOSE SAIRCRAFr, THE YTN-1T ( Y

Efficiency improved turboprop. Technical memo

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gearhart, W.S.

1982-06-10

Renewed attention has been focused on the efficiency of aircraft propulsion as the cost of fuel has risen. Studies conducted by NASA (1) to obtain fuel efficient aircraft have considered relatively highly-loaded turbo-prop systems. The disc loadings of these propellers are as much as four times higher than those on present turboprop aircraft. The higher disc loadings result in greater slipstream swirl and higher energy losses. Of primary importance is the radial distribution of the energy losses across the slipstream due to the tangential and axial velocities. This study presents the results of analysis defining the various sources of energymore » loss resulting from a swirling slipstream downstream of a propeller. Experimental data are presented demonstrating the presence of such losses and a propeller configuration discussed which offers improved propulsive performance when relatively highly-loaded propellers are employed.« less

Low-speed wind tunnel investigation of the static stability and control characteristics of an advanced turboprop configuration with the propellers placed over the tail. M.S. Thesis

NASA Technical Reports Server (NTRS)

Rhodes, Graham Scott

1990-01-01

An exploratory wind tunnel investigation was performed in the 30 x 60 foot wind tunnel to determine the low speed static stability and control characteristics into the deep stall regime of an advanced turboprop aircraft with the propellers located over the horizontal tail. By this arrangement, the horizontal tail could potentially provide acoustic shielding to reduce the high community noise caused by the propeller blades. The current configuration was a generic turboprop model equipped with 1 foot diameter single rotating eight bladed propellers that were designed for efficient cruise operation at a Mach number of 0.8. The data presented is static force data. The effects of power on the configuration characteristics were generally favorable. An arrangement with the propellers rotating with the outboard blades moving down was found to have significantly higher installed thrust than an arrangement with the propellers rotating with the inboard blades moving down. The primary unfavorable effect was a large pitch trim change which occurred with power, but the trim change could be minimized with a proper configuration design.

An experimental investigation of an advanced turboprop installation on a swept wing at subsonic and transonic speeds

NASA Technical Reports Server (NTRS)

Carlson, John R.; Pendergraft, Odis C., Jr.

1987-01-01

An investigation was conducted in the Langley 16-Foot Transonic Tunnel to determine the effects of a turboprop-nacelle installation on the pressure distributions over a swept, supercritical wing. The tests were conducted at Mach numbers from 0.20 to 0.80, at angles of attack from 0 to 5 degrees, nacelle nozzle pressure ratios from 1.0 to 1.6, and at propeller tip speeds from 700 to 800 ft/sec. The results of this study indicate that the turboprop nacelle interference, with and without power, on a swept wing is greater on the inboard wing panel than on the outboard wing panel. The over-the-wing nacelle installation with the propeller upwash on the inboard panel had flow separation problems at a Mach number of 0.80. No severe flow separation problems appear to exist for either propeller rotation direction for the under-the-wing nacelle installation. The local flow disturbances caused by the under-the-wing nacelle installation were in general less severe than for the over-the-wing nacelle installation.

General Electric TG-100A Turboprop in the Altitude Wind Tunnel

NASA Image and Video Library

1946-12-21

A General Electric TG-100A seen from the rear in the test section of the Altitude Wind Tunnel at the National Advisory Committee for Aeronautics (NACA) Lewis Flight Propulsion Laboratory in Cleveland, Ohio. The Altitude Wind Tunnel was used to study almost every model of US turbojet that emerged in the 1940s, as well as some ramjets and turboprops. In the early 1940s the military was interested in an engine that would use less fuel than the early jets but would keep up with them performance-wise. Turboprops seemed like a plausible solution. They could move a large volume of air and thus required less engine speed and less fuel. Researchers at General Electric’s plant in Schenectady, New York worked on the turboprop for several years in the 1930s. They received an army contract in 1941 to design a turboprop engine using an axial-flow compressor. The result was the 14-stage TG-100, the nation's first turboprop aircraft engine. Development of the engine was slow, however, and the military asked NACA Lewis to analyze the engine’s performance. The TG-100A was tested in the Altitude Wind Tunnel and it was determined that the compressors, combustion chamber, and turbine were impervious to changes in altitude. The researchers also established the optimal engine speed and propeller angle at simulated altitudes up to 35,000 feet. Despite these findings, development of the TG-100 was cancelled in May 1947. Twenty-eight of the engines were produced, but they were never incorporated into production aircraft.

Turboprop Propulsion Mechanic 2-8. Military Curriculum Materials for Vocational and Technical Education.

ERIC Educational Resources Information Center

Ohio State Univ., Columbus. National Center for Research in Vocational Education.

These military-developed curriculum materials for turboprop propulsion mechanics are targeted for use in grades 11-adult. Organized in five instructional blocks, the materials deal with the following topics: fundamentals of turboprop propulsion mechanics; engine and propeller systems operation; propeller maintenance; engine repair; and engine…

White matter tractography by means of Turboprop diffusion tensor imaging.

PubMed

Arfanakis, Konstantinos; Gui, Minzhi; Lazar, Mariana

2005-12-01

White matter fiber-tractography by means of diffusion tensor imaging (DTI) is a noninvasive technique that provides estimates of the structural connectivity of the brain. However, conventional fiber-tracking methods using DTI are based on echo-planar image acquisitions (EPI), which suffer from image distortions and artifacts due to magnetic susceptibility variations and eddy currents. Thus, a large percentage of white matter fiber bundles that are mapped using EPI-based DTI data are distorted, and/or terminated early, while others are completely undetected. This severely limits the potential of fiber-tracking techniques. In contrast, Turboprop imaging is a multiple-shot gradient and spin-echo (GRASE) technique that provides images with significantly fewer susceptibility and eddy current-related artifacts than EPI. The purpose of this work was to evaluate the performance of fiber-tractography techniques when using data obtained with Turboprop-DTI. All fiber pathways that were mapped were found to be in agreement with the anatomy. There were no visible distortions in any of the traced fiber bundles, even when these were located in the vicinity of significant magnetic field inhomogeneities. Additionally, the Turboprop-DTI data used in this research were acquired in less than 19 min of scan time. Thus, Turboprop appears to be a promising DTI data acquisition technique for tracing white matter fibers.

Simulation Modeling Requirements for Loss-of-Control Accident Prevention of Turboprop Transport Aircraft

NASA Technical Reports Server (NTRS)

Crider, Dennis; Foster, John V.

2012-01-01

In-flight loss of control remains the leading contributor to aviation accident fatalities, with stall upsets being the leading causal factor. The February 12, 2009. Colgan Air, Inc., Continental Express flight 3407 accident outside Buffalo, New York, brought this issue to the forefront of public consciousness and resulted in recommendations from the National Transportation Safety Board to conduct training that incorporates stalls that are fully developed and develop simulator standards to support such training. In 2010, Congress responded to this accident with Public Law 11-216 (Section 208), which mandates full stall training for Part 121 flight operations. Efforts are currently in progress to develop recommendations on implementation of stall training for airline pilots. The International Committee on Aviation Training in Extended Envelopes (ICATEE) is currently defining simulator fidelity standards that will be necessary for effective stall training. These recommendations will apply to all civil transport aircraft including straight-wing turboprop aircraft. Government-funded research over the previous decade provides a strong foundation for stall/post-stall simulation for swept-wing, conventional tail jets to respond to this mandate, but turboprops present additional and unique modeling challenges. First among these challenges is the effect of power, which can provide enhanced flow attachment behind the propellers. Furthermore, turboprops tend to operate for longer periods in an environment more susceptible to ice. As a result, there have been a significant number of turboprop accidents as a result of the early (lower angle of attack) stalls in icing. The vulnerability of turboprop configurations to icing has led to studies on ice accumulation and the resulting effects on flight behavior. Piloted simulations of these effects have highlighted the important training needs for recognition and mitigation of icing effects, including the reduction of stall margins

76 FR 45211 - Airworthiness Directives; Honeywell International Inc. TPE331-10 and TPE331-11 Series Turboprop...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-28

.... TPE331-10 and TPE331-11 Series Turboprop Engines AGENCY: Federal Aviation Administration, DOT. ACTION... and TPE331-11 series turboprop engines. That action would have required adding 360 first stage turbine... series turboprop engines, was published in the Federal Register on June 22, 2010 (75 FR 35354). The...

Piloted simulation study of an ILS approach of a twin-pusher business/commuter turboprop aircraft configuration

NASA Technical Reports Server (NTRS)

Riley, Donald R.; Brandon, Jay M.; Glaab, Louis J.

1994-01-01

A six-degree-of-freedom nonlinear simulation of a twin-pusher, turboprop business/commuter aircraft configuration representative of the Cessna ATPTB (Advanced turboprop test bed) was developed for use in piloted studies with the Langley General Aviation Simulator. The math models developed are provided, simulation predictions are compared with with Cessna flight-test data for validation purposes, and results of a handling quality study during simulated ILS (instrument landing system) approaches and missed approaches are presented. Simulated flight trajectories, task performance measures, and pilot evaluations are presented for the ILS approach and missed-approach tasks conducted with the vehicle in the presence of moderate turbulence, varying horizontal winds and engine-out conditions. Six test subjects consisting of two research pilots, a Cessna test pilot, and three general aviation pilots participated in the study. This effort was undertaken in cooperation with the Cessna Aircraft Company.

A Tractography Comparison between Turboprop and Spin-Echo Echo-Planar Diffusion Tensor Imaging

PubMed Central

Gui, Minzhi; Peng, Huiling; Carew, John D.; Lesniak, Maciej S.; Arfanakis, Konstantinos

2008-01-01

The development of accurate, non-invasive methods for mapping white matter fiber-tracts is of critical importance. However, fiber-tracking is typically performed on diffusion tensor imaging (DTI) data obtained with echo-planar-based imaging techniques (EPI), which suffer from susceptibility-related image artifacts, and image warping due to eddy-currents. Thus, a number of white matter fiber-bundles mapped using EPI-based DTI data are distorted and/or terminated early. This severely limits the clinical potential of fiber-tracking. In contrast, Turboprop-MRI provides images with significantly fewer susceptibility and eddy-current-related artifacts than EPI. The purpose of this work was to compare fiber-tracking results obtained from DTI data acquired with Turboprop-DTI and EPI-based DTI. It was shown that, in brain regions near magnetic field inhomogeneities, white matter fiber-bundles obtained with EPI-based DTI were distorted and/or partially detected, when magnetic susceptibility-induced distortions were not corrected. After correction, residual distortions were still present and several fiber-tracts remained partially detected. In contrast, when using Turboprop-DTI data, all traced fiber-tracts were in agreement with known anatomy. The inter-session reproducibility of tractography results was higher for Turboprop than EPI-based DTI data in regions near field inhomogeneities. Thus, Turboprop may be a more appropriate DTI data acquisition technique for tracing white matter fibers near regions with significant magnetic susceptibility differences, as well as in longitudinal studies of such fibers. However, the intra-session reproducibility of tractography results was higher for EPI-based than Turboprop DTI data. Thus, EPI-based DTI may be more advantageous for tracing fibers minimally affected by field inhomogeneities. PMID:18621131

A tractography comparison between turboprop and spin-echo echo-planar diffusion tensor imaging.

PubMed

Gui, Minzhi; Peng, Huiling; Carew, John D; Lesniak, Maciej S; Arfanakis, Konstantinos

2008-10-01

The development of accurate, non-invasive methods for mapping white matter fiber-tracts is of critical importance. However, fiber-tracking is typically performed on diffusion tensor imaging (DTI) data obtained with echo-planar-based imaging techniques (EPI), which suffer from susceptibility-related image artifacts, and image warping due to eddy-currents. Thus, a number of white matter fiber-bundles mapped using EPI-based DTI data are distorted and/or terminated early. This severely limits the clinical potential of fiber-tracking. In contrast, Turboprop-MRI provides images with significantly fewer susceptibility and eddy-current-related artifacts than EPI. The purpose of this work was to compare fiber-tracking results obtained from DTI data acquired with Turboprop-DTI and EPI-based DTI. It was shown that, in brain regions near magnetic field inhomogeneities, white matter fiber-bundles obtained with EPI-based DTI were distorted and/or partially detected, when magnetic susceptibility-induced distortions were not corrected. After correction, residual distortions were still present and several fiber-tracts remained partially detected. In contrast, when using Turboprop-DTI data, all traced fiber-tracts were in agreement with known anatomy. The inter-session reproducibility of tractography results was higher for Turboprop than EPI-based DTI data in regions near field inhomogeneities. Thus, Turboprop may be a more appropriate DTI data acquisition technique for tracing white matter fibers near regions with significant magnetic susceptibility differences, as well as in longitudinal studies of such fibers. However, the intra-session reproducibility of tractography results was higher for EPI-based than Turboprop DTI data. Thus, EPI-based DTI may be more advantageous for tracing fibers minimally affected by field inhomogeneities.

Quelling Cabin Noise in Turboprop Aircraft via Active Control

NASA Technical Reports Server (NTRS)

Kincaid, Rex K.; Laba, Keith E.; Padula, Sharon L.

1997-01-01

Cabin noise in turboprop aircraft causes passenger discomfort, airframe fatigue, and employee scheduling constraints due to OSHA standards for exposure to high levels of noise. The noise levels in the cabins of turboprop aircraft are typically 10 to 30 decibels louder than commercial jet noise levels. However. unlike jet noise the turboprop noise spectrum is dominated by a few low frequency tones. Active structural acoustic control is a method in which the control inputs (used to reduce interior noise) are applied directly to a vibrating structural acoustic system. The control concept modeled in this work is the application of in-plane force inputs to piezoceramic patches bonded to the wall of a vibrating cylinder. The goal is to determine the force inputs and locations for the piezoceramic actuators so that: (1) the interior noise is effectively damped; (2) the level of vibration of the cylinder shell is not increased; and (3) the power requirements needed to drive the actuators are not excessive. Computational experiments for data taken from a computer generated model and from a laboratory test article at NASA Langley Research Center are provided.

Analysis of in-flight acoustic data for a twin-engined turboprop airplane

NASA Technical Reports Server (NTRS)

Wilby, J. F.; Wilby, E. G.

1988-01-01

Acoustic measurements were made on the exterior and interior of a general aviation turboprop airplane during four flight tests. The test conditions were carefully controlled and repeated for each flight in order to determine data variability. For the first three flights the cabin was untreated and for the fourth flight the fuselage was treated with glass fiber batts. On the exterior, measured propeller harmonic sound pressure levels showed typical standard deviations of +1.4 dB, -2.3 dB, and turbulent boundary layer pressure levels, +1.2 dB, -1.6. Propeller harmonic levels in the cabin showed greater variability, with typical standard deviations of +2.0 dB, -4.2 dB. When interior sound pressure levels from different flights with different cabin treatments were used to evaluate insertion loss, the standard deviations were typically plus or minus 6.5 dB. This is due in part to the variability of the sound pressure level measurements, but probably is also influenced by changes in the model characteristics of the cabin. Recommendations are made for the planning and performance of future flight tests to measure interior noise of propeller-driven aircraft, either high-speed advanced turboprop or general aviation propellers.

Advanced Propfan Engine Technology (APET) and Single-rotation Gearbox/Pitch Change Mechanism

NASA Technical Reports Server (NTRS)

Sargisson, D. F.

1985-01-01

The projected performance, in the 1990's time period, of the equivalent technology level high bypass ratio turbofan powered aircraft (at the 150 passenger size) is compared with advanced turboprop propulsion systems. Fuel burn analysis, economic analysis, and pollution (noise, emissions) estimates were made. Three different cruise Mach numbers were investigated for both the turbofan and the turboprop systems. Aerodynamic design and performance estimates were made for nacelles, inlets, and exhaust systems. Air to oil heat exchangers were investigated for oil cooling advanced gearboxes at the 12,500 SHP level. The results and conclusions are positive in that high speed turboprop aircraft will exhibit superior fuel burn characteristics and lower operating costs when compared with equivalent technology turbofan aircraft.

Contribution of cardiac-induced brain pulsation to the noise of the diffusion tensor in Turboprop diffusion tensor imaging (DTI).

PubMed

Gui, Minzhi; Tamhane, Ashish A; Arfanakis, Konstantinos

2008-05-01

To assess the effects of cardiac-induced brain pulsation on the noise of the diffusion tensor in Turboprop (a form of periodically rotated overlapping parallel lines with enhanced reconstruction [PROPELLER] imaging) diffusion tensor imaging (DTI). A total of six healthy human subjects were imaged with cardiac-gated as well as nongated Turboprop DTI. Gated and nongated Turboprop DTI datasets were also simulated using actual data acquired exclusively during the diastolic or systolic period of the cardiac cycle. The total variance of the diffusion tensor (TVDT) was measured and compared between acquisitions. The TVDT near the ventricles was significantly reduced in cardiac-gated compared to nongated Turboprop DTI acquisitions. Furthermore, the effects of brain pulsation were reduced, but not eliminated, when increasing the amount of data collected. Finally, data corrupted by cardiac-induced pulsation were not consistently detected by the step of the conventional Turboprop reconstruction algorithm that evaluates the quality of data in different blades. Thus, the inherent quality weighting of the conventional Turboprop reconstruction algorithm was unable to compensate for the increased noise in the diffusion tensor due to brain pulsation. Cardiac-induced brain pulsation increases the TVDT in Turboprop DTI. Use of cardiac gating to limit data acquisition to the diastolic period of the cardiac cycle reduces the TVDT at the expense of imaging time. (c) 2008 Wiley-Liss, Inc.

Motion correction in periodically-rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) and turboprop MRI.

PubMed

Tamhane, Ashish A; Arfanakis, Konstantinos

2009-07-01

Periodically-rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) and Turboprop MRI are characterized by greatly reduced sensitivity to motion, compared to their predecessors, fast spin-echo (FSE) and gradient and spin-echo (GRASE), respectively. This is due to the inherent self-navigation and motion correction of PROPELLER-based techniques. However, it is unknown how various acquisition parameters that determine k-space sampling affect the accuracy of motion correction in PROPELLER and Turboprop MRI. The goal of this work was to evaluate the accuracy of motion correction in both techniques, to identify an optimal rotation correction approach, and determine acquisition strategies for optimal motion correction. It was demonstrated that blades with multiple lines allow more accurate estimation of motion than blades with fewer lines. Also, it was shown that Turboprop MRI is less sensitive to motion than PROPELLER. Furthermore, it was demonstrated that the number of blades does not significantly affect motion correction. Finally, clinically appropriate acquisition strategies that optimize motion correction are discussed for PROPELLER and Turboprop MRI. (c) 2009 Wiley-Liss, Inc.

Artificial intelligence for turboprop engine maintenance

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1995-01-01

Long-term maintenance operations, causing the unit to out of action, may seem economical - but they result in reduced operating readiness. Offsetting that concern, careless, hurried maintenance reduces margins of safety and reliability. Any tool that improves maintenance without causing a sharp increase in cost is valuable. Artificial intelligence (AI) is one of the tools. Expert system and neural networks are two different areas of AI that show promise for turboprop engine maintenance.

Turboprop: improved PROPELLER imaging.

PubMed

Pipe, James G; Zwart, Nicholas

2006-02-01

A variant of periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) MRI, called turboprop, is introduced. This method employs an oscillating readout gradient during each spin echo of the echo train to collect more lines of data per echo train, which reduces the minimum scan time, motion-related artifact, and specific absorption rate (SAR) while increasing sampling efficiency. It can be applied to conventional fast spin-echo (FSE) imaging; however, this article emphasizes its application in diffusion-weighted imaging (DWI). The method is described and compared with conventional PROPELLER imaging, and clinical images collected with this PROPELLER variant are shown. Copyright 2006 Wiley-Liss, Inc.

Advanced turboprop testbed systems study

NASA Technical Reports Server (NTRS)

Goldsmith, I. M.

1982-01-01

The proof of concept, feasibility, and verification of the advanced prop fan and of the integrated advanced prop fan aircraft are established. The use of existing hardware is compatible with having a successfully expedited testbed ready for flight. A prop fan testbed aircraft is definitely feasible and necessary for verification of prop fan/prop fan aircraft integrity. The Allison T701 is most suitable as a propulsor and modification of existing engine and propeller controls are adequate for the testbed. The airframer is considered the logical overall systems integrator of the testbed program.

Effects of vibration on the readability of an electronic flight instrument display

NASA Astrophysics Data System (ADS)

Viveash, Jacqueline P.; Cable, A. N.; King, S. K.; Stott, J. R.; Wright, R.

1993-12-01

An in-flight icing incident involving a BAe advanced turboprop (ATP) aircraft led to severe vibration of the airframe and a loss of aerodynamic control. During the period of vibration the pilot reported a specific pattern of image break up on the electronic flight instrument system (EFIS). Three experiments to investigate this visual effect are reported.

75 FR 35354 - Airworthiness Directives; Honeywell International Inc. TPE331-10 and TPE331-11 Series Turboprop...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-22

... International Inc. TPE331-10 and TPE331-11 Series Turboprop Engines AGENCY: Federal Aviation Administration (FAA... airworthiness directive (AD) for Honeywell International Inc. TPE331-10 and TPE331-11 series turboprop engines... likely to exist or develop on other products of this same type design. For that reason, we are proposing...

Turboprop engine and method of operating the same

DOE Office of Scientific and Technical Information (OSTI.GOV)

Klees, G.W.; Johnson, P.E.

1986-02-11

This patent describes a turboprop engine consisting of: 1.) A compressor; 2.) A turbine; 3.) A combustion section; 4.) A variable pitch propeller; 5.) A speed reducing transmission; 6.) An air inlet; 7.) An air inlet bypass; 8.) An air outlet bypass duct; 9.) A flow control operatively positioned to receive air flow from the air inlet bypass and air flow from the low pressure compressor component. To direct the air flow to the air outlet bypass duct, and the air flow to the high pressure compressor component, the flow control has a first position where the air flow ismore » from. The high and low pressure compressor components and is directed to the air outlet bypass duct. The flow control has a second position for the air flow from the air inlet bypass duct to the air outlet bypass duct and air from the low pressure compressor component is directed to the high pressure compressor component. A method of operating a turboprop engine.« less

75 FR 51187 - Airworthiness Directives; Pratt & Whitney Canada Corp. (P&WC) PW305A and PW305B Turboprop Engines

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-19

... Airworthiness Directives; Pratt & Whitney Canada Corp. (P&WC) PW305A and PW305B Turboprop Engines AGENCY... and PW305B turboprop engines with certain impellers, part numbers (P/Ns) 30B2185, 30B2486, 30B2858-01...

Aeroelastic stability and response of rotating structures

NASA Technical Reports Server (NTRS)

Keith, Theo G., Jr.

1993-01-01

A summary of the work performed during the progress period is presented. Analysis methods for predicting loads and instabilities of wind turbines were developed. Three new areas of research to aid the Advanced Turboprop Project (ATP) were initiated and developed. These three areas of research are aeroelastic analysis methods for cascades including blade and disk flexibility; stall flutter analysis; and computational aeroelasticity.

Python Turboprop Prepared for a Test in the Altitude Wind Tunnel

NASA Image and Video Library

1949-08-21

A 3670-horsepower Armstrong-Siddeley Python turboprop being prepared for tests in the Altitude Wind Tunnel at the National Advisory Committee for Aeronautics (NACA) Lewis Flight Propulsion Laboratory. In 1947 Lewis researcher Walter Olsen led a group of representatives from the military, industry, and the NACA on a fact finding mission to investigate the technological progress of British turbojet manufacturers. Afterwards several British engines, including the Python, were brought to Cleveland for testing in Lewis’s altitude facilities. The Python was a 14-stage axial-flow compressor turboprop with a fixed-area nozzle and contra-rotating propellers. Early turboprops combined the turbojet and piston engine technologies. They could move large quantities of air so required less engine speed and thus less fuel. This was very appealing to the military for some applications. The military asked the NACA to compare the Python’s performance at sea to that at high altitudes. The NACA researchers studied the Python in the Altitude Wind Tunnel from July 1949 through January 1950. It was the first time the tunnel was used to study an engine with the sole purpose of learning about, not improving it. They analyzed the engine’s dynamic response using a frequency response method at altitudes between 10,000 to 30,000 feet. Lewis researchers found that they could predict the dynamic response characteristics at any altitude from the data obtained from any other specific altitude. This portion of the testing was completed during a single test run.

Performance estimation for a highly loaded eight-blade propeller combined with an advanced technology turboshaft engine

NASA Technical Reports Server (NTRS)

Morris, S. J., Jr.

1979-01-01

Performance estimation, weights, and scaling laws for an eight-blade highly loaded propeller combined with an advanced turboshaft engine are presented. The data are useful for planned aircraft mission studies using the turboprop propulsion system. Comparisons are made between the performance of the 1990+ technology turboprop propulsion system and the performance of both a current technology turbofan and an 1990+ technology turbofan.

Research on Modeling of Propeller in a Turboprop Engine

NASA Astrophysics Data System (ADS)

Huang, Jiaqin; Huang, Xianghua; Zhang, Tianhong

2015-05-01

In the simulation of engine-propeller integrated control system for a turboprop aircraft, a real-time propeller model with high-accuracy is required. A study is conducted to compare the real-time and precision performance of propeller models based on strip theory and lifting surface theory. The emphasis in modeling by strip theory is focused on three points as follows: First, FLUENT is adopted to calculate the lift and drag coefficients of the propeller. Next, a method to calculate the induced velocity which occurs in the ground rig test is presented. Finally, an approximate method is proposed to obtain the downwash angle of the propeller when the conventional algorithm has no solution. An advanced approximation of the velocities induced by helical horseshoe vortices is applied in the model based on lifting surface theory. This approximate method will reduce computing time and remain good accuracy. Comparison between the two modeling techniques shows that the model based on strip theory which owns more advantage on both real-time and high-accuracy can meet the requirement.

Cost/benefit analysis of advanced materials technology candidates for the 1980's, part 2

NASA Technical Reports Server (NTRS)

Dennis, R. E.; Maertins, H. F.

1980-01-01

Cost/benefit analyses to evaluate advanced material technologies projects considered for general aviation and turboprop commuter aircraft through estimated life-cycle costs, direct operating costs, and development costs are discussed. Specifically addressed is the selection of technologies to be evaluated; development of property goals; assessment of candidate technologies on typical engines and aircraft; sensitivity analysis of the changes in property goals on performance and economics, cost, and risk analysis for each technology; and ranking of each technology by relative value. The cost/benefit analysis was applied to a domestic, nonrevenue producing, business-type jet aircraft configured with two TFE731-3 turbofan engines, and to a domestic, nonrevenue producing, business type turboprop aircraft configured with two TPE331-10 turboprop engines. In addition, a cost/benefit analysis was applied to a commercial turboprop aircraft configured with a growth version of the TPE331-10.

Escape strategies for turboprop aircraft in microburst windshear

NASA Technical Reports Server (NTRS)

Bobbitt, Richard B.; Howard, Richard M.

1991-01-01

The dynamic reponse of a P-3 aircraft and a light twin-engine turboprop to a low-level microburst encounter is modeled. The response to the microburst is depicted for various escape maneuvers. Plots of altitude, velocity, and specific energy are shown for all cases. Takeoff escape strategies are discussed. The optimal escape procedure is found to be flying a constant value of pitch angle. Constant angle of attack maneuvers sometimes result in superior performance.

Turboprop IDEAL: a motion-resistant fat-water separation technique.

PubMed

Huo, Donglai; Li, Zhiqiang; Aboussouan, Eric; Karis, John P; Pipe, James G

2009-01-01

Suppression of the fat signal in MRI is very important for many clinical applications. Multi-point water-fat separation methods, such as IDEAL (Iterative Decomposition of water and fat with Echo Asymmetry and Least-squares estimation), can robustly separate water and fat signal, but inevitably increase scan time, making separated images more easily affected by patient motions. PROPELLER (Periodically Rotated Overlapping ParallEL Lines with Enhanced Reconstruction) and Turboprop techniques offer an effective approach to correct for motion artifacts. By combining these techniques together, we demonstrate that the new TP-IDEAL method can provide reliable water-fat separation with robust motion correction. The Turboprop sequence was modified to acquire source images, and motion correction algorithms were adjusted to assure the registration between different echo images. Theoretical calculations were performed to predict the optimal shift and spacing of the gradient echoes. Phantom images were acquired, and results were compared with regular FSE-IDEAL. Both T1- and T2-weighted images of the human brain were used to demonstrate the effectiveness of motion correction. TP-IDEAL images were also acquired for pelvis, knee, and foot, showing great potential of this technique for general clinical applications.

High speed turboprop aeroacoustic study (counterrotation). Volume 2: Computer programs

NASA Technical Reports Server (NTRS)

Whitfield, C. E.; Mani, R.; Gliebe, P. R.

1990-01-01

The isolated counterrotating high speed turboprop noise prediction program developed and funded by GE Aircraft Engines was compared with model data taken in the GE Aircraft Engines Cell 41 anechoic facility, the Boeing Transonic Wind Tunnel, and in the NASA-Lewis 8 x 6 and 9 x 15 wind tunnels. The predictions show good agreement with measured data under both low and high speed simulated flight conditions. The installation effect model developed for single rotation, high speed turboprops was extended to include counter rotation. The additional effect of mounting a pylon upstream of the forward rotor was included in the flow field modeling. A nontraditional mechanism concerning the acoustic radiation from a propeller at angle of attack was investigated. Predictions made using this approach show results that are in much closer agreement with measurement over a range of operating conditions than those obtained via traditional fluctuating force methods. The isolated rotors and installation effects models were combined into a single prediction program. The results were compared with data taken during the flight test of the B727/UDF (trademark) engine demonstrator aircraft.

High speed turboprop aeroacoustic study (counterrotation). Volume 2: Computer programs

NASA Astrophysics Data System (ADS)

Whitfield, C. E.; Mani, R.; Gliebe, P. R.

1990-07-01

The isolated counterrotating high speed turboprop noise prediction program developed and funded by GE Aircraft Engines was compared with model data taken in the GE Aircraft Engines Cell 41 anechoic facility, the Boeing Transonic Wind Tunnel, and in the NASA-Lewis 8 x 6 and 9 x 15 wind tunnels. The predictions show good agreement with measured data under both low and high speed simulated flight conditions. The installation effect model developed for single rotation, high speed turboprops was extended to include counter rotation. The additional effect of mounting a pylon upstream of the forward rotor was included in the flow field modeling. A nontraditional mechanism concerning the acoustic radiation from a propeller at angle of attack was investigated. Predictions made using this approach show results that are in much closer agreement with measurement over a range of operating conditions than those obtained via traditional fluctuating force methods. The isolated rotors and installation effects models were combined into a single prediction program. The results were compared with data taken during the flight test of the B727/UDF (trademark) engine demonstrator aircraft.

Performance estimation for highly loaded six and ten blade propellers combined with an advanced technology turboshaft engine

NASA Technical Reports Server (NTRS)

Morris, S. J., Jr.

1980-01-01

Performance estimations, weights, and scaling laws for the six blade and ten blade highly loaded propellers combined with an advanced turboshaft engine are presented. These data are useful for aircraft mission studies using the turboprop system. Comparisons are made between the performance of post 1980 technology turboprop propulsion systems and the performance of both a current technology turbofan and a post 1990 technology turbofan.

Aeropropulsion 1987. Session 5: Subsonic Propulsion Technology

NASA Technical Reports Server (NTRS)

1987-01-01

NASA is conducting aeropropulsion research over a broad range of Mach numbers. In addition to the high-speed propulsion research described, major progress was recorded in research aimed at the subsonic flight regimes of interest to many commercial and military users. Recent progress and future directions in such areas as small engine technology, rotorcraft transmissions, icing, Hot Section Technology (HOST) and the Advanced Turboprop Program (ATP) are covered.

Aeropropulsion '87. Session 5: Subsonic propulsion technology

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1987-11-01

NASA is conducting aeropropulsion research over a broad range of Mach numbers. In addition to the high-speed propulsion research described, major progress was recorded in research aimed at the subsonic flight regimes of interest to many commercial and military users. Recent progress and future directions in such areas as small engine technology, rotorcraft transmissions, icing, Hot Section Technology (HOST) and the Advanced Turboprop Program (ATP) are covered.

Performance of a Turboprop Engine with Heat Recovery in Off-Design Conditions

NASA Astrophysics Data System (ADS)

Andriani, Roberto; Ghezzi, Umberto; Gamma, Fausto; Ingenito, Antonella; Agresta, Antonio

2013-09-01

The research for fuel consumption and pollution reduction in new generation aero engines has indicated intercooling and regeneration as very effective methods for this purpose. Hence, different countries have joined their efforts in common research programs, to develop new gas turbine engines able to reduce considerably the fuel consumption and the ambient impact by means of these two techniques. To study their effects on the engine performance and characteristics, a thermodynamic numerical program that simulates the behavior of a turboprop engine with intercooling and regeneration in different operating conditions has been developed. After the parametric study, and the definition of the design conditions, the off-design analysis is carried on, comparing the main characteristics of the intercooled-regenerated turboprop with those of a conventional engine. Then, once a particular mission profile was fixed, the engine performance, in particular the equivalent power, the fuel consumption and the heat exchanger weight were discussed.

Fuel Consumption Reduction and Weight Estimate of an Intercooled-Recuperated Turboprop Engine

NASA Astrophysics Data System (ADS)

Andriani, Roberto; Ghezzi, Umberto; Ingenito, Antonella; Gamma, Fausto

2012-09-01

The introduction of intercooling and regeneration in a gas turbine engine can lead to performance improvement and fuel consumption reduction. Moreover, as first consequence of the saved fuel, also the pollutant emission can be greatly reduced. Turboprop seems to be the most suitable gas turbine engine to be equipped with intercooler and heat recuperator thanks to the relatively small mass flow rate and the small propulsion power fraction due to the exhaust nozzle. However, the extra weight and drag due to the heat exchangers must be carefully considered. An intercooled-recuperated turboprop engine is studied by means of a thermodynamic numeric code that, computing the thermal cycle, simulates the engine behavior at different operating conditions. The main aero engine performances, as specific power and specific fuel consumption, are then evaluated from the cycle analysis. The saved fuel, the pollution reduction, and the engine weight are then estimated for an example case.

Exergo-Economic Analysis of an Experimental Aircraft Turboprop Engine Under Low Torque Condition

NASA Astrophysics Data System (ADS)

Atilgan, Ramazan; Turan, Onder; Aydin, Hakan

Exergo-economic analysis is an unique combination of exergy analysis and cost analysis conducted at the component level. In exergo-economic analysis, cost of each exergy stream is determined. Inlet and outlet exergy streams of the each component are associated to a monetary cost. This is essential to detect cost-ineffective processes and identify technical options which could improve the cost effectiveness of the overall energy system. In this study, exergo-economic analysis is applied to an aircraft turboprop engine. Analysis is based on experimental values at low torque condition (240 N m). Main components of investigated turboprop engine are the compressor, the combustor, the gas generator turbine, the free power turbine and the exhaust. Cost balance equations have been formed for all components individually and exergo-economic parameters including cost rates and unit exergy costs have been calculated for each component.

An Investigation of Landing-Contact Conditions for Two Large Turbojet Transports and a Turboprop Transport During Routine Daylight Operations

NASA Technical Reports Server (NTRS)

Stickle, Joseph W.

1961-01-01

The National Aeronautics and Space Administration has recently completed a statistical investigation of landing-contact conditions for two large turbojet transports and a turboprop transport landing on a dry runway during routine daylight operations at the Los Angeles International Airport. Measurements were made to obtain vertical velocity, airspeed, rolling velocity, bank angle, and distance from the runway threshold, just prior to ground contact. The vertical velocities at touchdown for one of the turbojet airplanes measured in this investigation were essentially the same as those measured on the same type of airplane during a similar investigation (see NASA Technical Note D-527) conducted approximately 8 months earlier. Thus, it appeared that 8 months of additional pilot experience has had no noticeable tendency toward lowering the vertical velocities of this transport. Distributions of vertical velocities for the turbojet transports covered in this investigation were similar and considerably higher than'those for the turboprop transport. The data for the turboprop transport were in good agreement with the data for the piston-engine transports (see NACA Report 1214 and NASA Technical Note D-147) for all the measured parameters. For the turbojet transports, 1 landing in 100 would be expected to equal or exceed a vertical velocity of approximately 4.2 ft/sec; whereas, for the turboprop transport, 1 landing in 100 would be expected to equal or exceed 3.2 ft/sec. The mean airspeeds at touchdown for the three transports ranged from 22.5 percent to 26.6 percent above the stalling speed. Rolling velocities for the turbojet transports were considerably higher than those for the turboprop transport. Distributions of bank angles at contact for the three transports were similar. For each type of airplane, 1 landing in 100 would be expected to equal or exceed a bank angle at touchdown of approximately 3.0 deg. Distributions of touchdown distances for the three transports

PERFORMANCE OF TWO LIQUID METAL TURBOPROP ENGINES UTILIZING A CIRCULATING FUEL REACTOR

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tiedemann, H.J.; Mathews, L.

1955-01-20

The performance of two all-nuclear turboprop engines utilizing the circulating fuel reactor with a fluoride fuel temperature of I500 deg F was investigated. Data are presented for off-match-point and modified match-point performances. Results are given in graph form. (M.C.G.)

AIRCRAFT REACTOR CONTROL SYSTEM APPLICABLE TO TURBOJET AND TURBOPROP POWER PLANTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gorker, G.E.

1955-07-19

Control systems proposed for direct cycle nuclear powered aircraft commonly involve control of engine speed, nuclear energy input, and chcmical energy input. A system in which these parameters are controlled by controlling the total energy input, the ratio of nuclear and chemical energy input, and the engine speed is proposed. The system is equally applicable to turbojet or turboprop applications. (auth)

Application of advanced technologies to small, short-haul transport aircraft

NASA Technical Reports Server (NTRS)

Coussens, T. G.; Tullis, R. H.

1980-01-01

The performance and economic benefits available by incorporation of advanced technologies into the small, short haul air transport were assessed. Low cost structure and advanced composite material, advanced turboprop engines and new propellers, advanced high lift systems and active controls; and alternate aircraft configurations with aft mounted engines were investigated. Improvements in fuel consumed and aircraft economics (acquisition cost and direct operating cost) are available by incorporating selected advanced technologies into the small, short haul aircraft.

High speed turboprop aeroacoustic study (counterrotation). Volume 1: Model development

NASA Technical Reports Server (NTRS)

Whitfield, C. E.; Mani, R.; Gliebe, P. R.

1990-01-01

The isolated counterrotating high speed turboprop noise prediction program was compared with model data taken in the GE Aircraft Engines Cell 41 anechoic facility, the Boeing Transonic Wind Tunnel, and in NASA-Lewis' 8x6 and 9x15 wind tunnels. The predictions show good agreement with measured data under both low and high speed simulated flight conditions. The installation effect model developed for single rotation, high speed turboprops was extended to include counterotation. The additional effect of mounting a pylon upstream of the forward rotor was included in the flow field modeling. A nontraditional mechanism concerning the acoustic radiation from a propeller at angle of attach was investigated. Predictions made using this approach show results that are in much closer agreement with measurement over a range of operating conditions than those obtained via traditional fluctuating force methods. The isolated rotors and installation effects models were combines into a single prediction program, results of which were compared with data taken during the flight test of the B727/UDF engine demonstrator aircraft. Satisfactory comparisons between prediction and measured data for the demonstrator airplane, together with the identification of a nontraditional radiation mechanism for propellers at angle of attack are achieved.

High speed turboprop aeroacoustic study (counterrotation). Volume 1: Model development

NASA Astrophysics Data System (ADS)

Whitfield, C. E.; Mani, R.; Gliebe, P. R.

1990-07-01

The isolated counterrotating high speed turboprop noise prediction program was compared with model data taken in the GE Aircraft Engines Cell 41 anechoic facility, the Boeing Transonic Wind Tunnel, and in NASA-Lewis' 8x6 and 9x15 wind tunnels. The predictions show good agreement with measured data under both low and high speed simulated flight conditions. The installation effect model developed for single rotation, high speed turboprops was extended to include counterotation. The additional effect of mounting a pylon upstream of the forward rotor was included in the flow field modeling. A nontraditional mechanism concerning the acoustic radiation from a propeller at angle of attach was investigated. Predictions made using this approach show results that are in much closer agreement with measurement over a range of operating conditions than those obtained via traditional fluctuating force methods. The isolated rotors and installation effects models were combines into a single prediction program, results of which were compared with data taken during the flight test of the B727/UDF engine demonstrator aircraft. Satisfactory comparisons between prediction and measured data for the demonstrator airplane, together with the identification of a nontraditional radiation mechanism for propellers at angle of attack are achieved.

Application of advanced technologies to small, short-haul aircraft

NASA Technical Reports Server (NTRS)

Andrews, D. G.; Brubaker, P. W.; Bryant, S. L.; Clay, C. W.; Giridharadas, B.; Hamamoto, M.; Kelly, T. J.; Proctor, D. K.; Myron, C. E.; Sullivan, R. L.

1978-01-01

The results of a preliminary design study which investigates the use of selected advanced technologies to achieve low cost design for small (50-passenger), short haul (50 to 1000 mile) transports are reported. The largest single item in the cost of manufacturing an airplane of this type is labor. A careful examination of advanced technology to airframe structure was performed since one of the most labor-intensive parts of the airplane is structures. Also, preliminary investigation of advanced aerodynamics flight controls, ride control and gust load alleviation systems, aircraft systems and turbo-prop propulsion systems was performed. The most beneficial advanced technology examined was bonded aluminum primary structure. The use of this structure in large wing panels and body sections resulted in a greatly reduced number of parts and fasteners and therefore, labor hours. The resultant cost of assembled airplane structure was reduced by 40% and the total airplane manufacturing cost by 16% - a major cost reduction. With further development, test verification and optimization appreciable weight saving is also achievable. Other advanced technology items which showed significant gains are as follows: (1) advanced turboprop-reduced block fuel by 15.30% depending on range; (2) configuration revisions (vee-tail)-empennage cost reduction of 25%; (3) leading-edge flap addition-weight reduction of 2500 pounds.

Genomic Analysis of ATP Efflux in Saccharomyces cerevisiae

PubMed Central

Peters, Theodore W.; Miller, Aaron W.; Tourette, Cendrine; Agren, Hannah; Hubbard, Alan; Hughes, Robert E.

2015-01-01

Adenosine triphosphate (ATP) plays an important role as a primary molecule for the transfer of chemical energy to drive biological processes. ATP also functions as an extracellular signaling molecule in a diverse array of eukaryotic taxa in a conserved process known as purinergic signaling. Given the important roles of extracellular ATP in cell signaling, we sought to comprehensively elucidate the pathways and mechanisms governing ATP efflux from eukaryotic cells. Here, we present results of a genomic analysis of ATP efflux from Saccharomyces cerevisiae by measuring extracellular ATP levels in cultures of 4609 deletion mutants. This screen revealed key cellular processes that regulate extracellular ATP levels, including mitochondrial translation and vesicle sorting in the late endosome, indicating that ATP production and transport through vesicles are required for efflux. We also observed evidence for altered ATP efflux in strains deleted for genes involved in amino acid signaling, and mitochondrial retrograde signaling. Based on these results, we propose a model in which the retrograde signaling pathway potentiates amino acid signaling to promote mitochondrial respiration. This study advances our understanding of the mechanism of ATP secretion in eukaryotes and implicates TOR complex 1 (TORC1) and nutrient signaling pathways in the regulation of ATP efflux. These results will facilitate analysis of ATP efflux mechanisms in higher eukaryotes. PMID:26585826

ATP: A Coherent View for School Advanced Level Studies in Biology.

ERIC Educational Resources Information Center

Gayford, Chris

1986-01-01

Discusses how instruction of biological concepts as ATP cellular energetics is related to fundamental physical science understandings. Reviews areas of common misconceptions and confusions. Summarizes results of a study which investigated students' knowledge and perception of difficulty associated with the topic of energy and ATP. (ML)

Aerodynamic design and performance testing of an advanced 30 deg swept, eight bladed propeller at Mach numbers from 0.2 to 0.85

NASA Technical Reports Server (NTRS)

Black, D. M.; Menthe, R. W.; Wainauski, H. S.

1978-01-01

The increased emphasis on fuel conservation in the world has stimulated a series of studies of both conventional and unconventional propulsion systems for commercial aircraft. Preliminary results from these studies indicate that a fuel saving of from 15 to 28 percent may be realized by the use of an advanced high speed turboprop. The turboprop must be capable of high efficiency at Mach 0.8 above 10.68 km (35,000 ft) altitude if it is to compete with turbofan powered commercial aircraft. An advanced turboprop concept was wind tunnel tested. The model included such concepts as an aerodynamically integrated propeller/nacelle, blade sweep and power (disk) loadings approximately three times higher than conventional propeller designs. The aerodynamic design for the model is discussed. Test results are presented which indicate propeller net efficiencies near 80 percent were obtained at high disk loadings at Mach 0.8.

Optimization of a Turboprop UAV for Maximum Loiter and Specific Power Using Genetic Algorithm

NASA Astrophysics Data System (ADS)

Dinc, Ali

2016-09-01

In this study, a genuine code was developed for optimization of selected parameters of a turboprop engine for an unmanned aerial vehicle (UAV) by employing elitist genetic algorithm. First, preliminary sizing of a UAV and its turboprop engine was done, by the code in a given mission profile. Secondly, single and multi-objective optimization were done for selected engine parameters to maximize loiter duration of UAV or specific power of engine or both. In single objective optimization, as first case, UAV loiter time was improved with an increase of 17.5% from baseline in given boundaries or constraints of compressor pressure ratio and burner exit temperature. In second case, specific power was enhanced by 12.3% from baseline. In multi-objective optimization case, where previous two objectives are considered together, loiter time and specific power were increased by 14.2% and 9.7% from baseline respectively, for the same constraints.

Aerodynamic design and performance testing of an advanced 30 deg swept, eight bladed propeller at Mach numbers from 0. 2 to 0. 85. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Black, D.M.; Menthe, R.W.; Wainauski, H.S.

1978-09-01

The increased emphasis on fuel conservation in the world has stimulated a series of studies of both conventional and unconventional propulsion systems for commercial aircraft. Preliminary results from these studies indicate that a fuel saving of from 15 to 28 percent may be realized by the use of an advanced high speed turboprop. The turboprop must be capable of high efficiency at Mach 0.8 above 10.68 km (35,000 ft) altitude if it is to compete with turbofan powered commercial aircraft. An advanced turboprop concept was wind tunnel tested. The model included such concepts as an aerodynamically integrated propeller/nacelle, blade sweepmore » and power (disk) loadings approximately three times higher than conventional propeller designs. The aerodynamic design for the model is discussed. Test results are presented which indicate propeller net efficiencies near 80 percent were obtained at high disk loadings at Mach 0.8.« less

Customized ATP towpreg. [Automated Tow Placement

NASA Technical Reports Server (NTRS)

Sandusky, Donald A.; Marchello, Joseph M.; Baucom, Robert M.; Johnston, Norman J.

1992-01-01

Automated tow placement (ATP) utilizes robotic technology to lay down adjacent polymer-matrix-impregnated carbon fiber tows on a tool surface. Consolidation and cure during ATP requires that void elimination and polymer matrix adhesion be accomplished in the short period of heating and pressure rolling that follows towpreg ribbon placement from the robot head to the tool. This study examined the key towpreg ribbon properties and dimensions which play a significant role in ATP. Analysis of the heat transfer process window indicates that adequate heating can be achieved at lay down rates as high as 1 m/sec. While heat transfer did not appear to be the limiting factor, resin flow and fiber movement into tow lap gaps could be. Accordingly, consideration was given to towpreg ribbon having uniform yet non-rectangular cross sections. Dimensional integrity of the towpreg ribbon combined with customized ribbon architecture offer great promise for processing advances in ATP of high performance composites.

ATP Synthase, a Target for Dementia and Aging?

PubMed

Larrick, James W; Larrick, Jasmine W; Mendelsohn, Andrew R

2018-02-01

Advancing age is the biggest risk factor for development for the major life-threatening diseases in industrialized nations accounting for >90% of deaths. Alzheimer's dementia (AD) is among the most devastating. Currently approved therapies fail to slow progression of the disease, providing only modest improvements in memory. Recently reported work describes mechanistic studies of J147, a promising therapeutic molecule previously shown to rescue the severe cognitive deficits exhibited by aged, transgenic AD mice. Apparently, J147 targets the mitochondrial alpha-F1-ATP synthase (ATP5A). Modest inhibition of the ATP synthase modulates intracellular calcium to activate AMP-activated protein kinase to inhibit mammalian target of rapamycin, a known mechanism of lifespan extension from worms to mammals.

Advanced aerodynamics. Selected NASA research

NASA Technical Reports Server (NTRS)

1981-01-01

This Conference Publication contains selected NASA papers that were presented at the Fifth Annual Status Review of the NASA Aircraft Energy Efficiency (ACEE) Energy Efficient Transport (EET) Program held at Dryden Flight Research Center in Edwards, California on September 14 to 15, 1981. These papers describe the status of several NASA in-house research activities in the areas of advanced turboprops, natural laminar flow, oscillating control surfaces, high-Reynolds-number airfoil tests, high-lift technology, and theoretical design techniques.

The Chloroplast ATP Synthase Features the Characteristic Redox Regulation Machinery

PubMed Central

Sunamura, Ei-Ichiro; Kim, Yusung; Konno, Hiroki

2013-01-01

Abstract Significance: Regulation of the activity of the chloroplast ATP synthase is largely accomplished by the chloroplast thioredoxin system, the main redox regulation system in chloroplasts, which is directly coupled to the photosynthetic reaction. We review the current understanding of the redox regulation system of the chloroplast ATP synthase. Recent Advances: The thioredoxin-targeted portion of the ATP synthase consists of two cysteines located on the central axis subunit γ. The redox state of these two cysteines is under the influence of chloroplast thioredoxin, which directly controls rotation during catalysis by inducing a conformational change in this subunit. The molecular mechanism of redox regulation of the chloroplast ATP synthase has recently been determined. Critical Issues: Regulation of the activity of the chloroplast ATP synthase is critical in driving efficiency into the ATP synthesis reaction in chloroplasts. Future Directions: The molecular architecture of the chloroplast ATP synthase, which confers redox regulatory properties requires further investigation, in light of the molecular structure of the enzyme complex as well as the physiological significance of the regulation system. Antioxid. Redox Signal. 19, 1846–1854. PMID:23145525

Unified Aeroacoustics Analysis for High Speed Turboprop Aerodynamics and Noise. Volume 1; Development of Theory for Blade Loading, Wakes, and Noise

NASA Technical Reports Server (NTRS)

Hanson, D. B.

1991-01-01

A unified theory for the aerodynamics and noise of advanced turboprops are presented. Aerodynamic topics include calculation of performance, blade load distribution, and non-uniform wake flow fields. Blade loading can be steady or unsteady due to fixed distortion, counter-rotating wakes, or blade vibration. The aerodynamic theory is based on the pressure potential method and is therefore basically linear. However, nonlinear effects associated with finite axial induction and blade vortex flow are included via approximate methods. Acoustic topics include radiation of noise caused by blade thickness, steady loading (including vortex lift), and unsteady loading. Shielding of the fuselage by its boundary layer and the wing are treated in separate analyses that are compatible but not integrated with the aeroacoustic theory for rotating blades.

Noise transmission and reduction in turboprop aircraft

NASA Astrophysics Data System (ADS)

MacMartin, Douglas G.; Basso, Gordon L.; Leigh, Barry

1994-09-01

There is considerable interest in reducing the cabin noise environment in turboprop aircraft. Various approaches have been considered at deHaviland Inc., including passive tuned-vibration absorbers, speaker-based noise cancellation, and structural vibration control of the fuselage. These approaches will be discussed briefly. In addition to controlling the noise, a method of predicting the internal noise is required both to evaluate potential noise reduction approaches, and to validate analytical design models. Instead of costly flight tests, or carrying out a ground simulation of the propeller pressure field, a much simpler reciprocal technique can be used. A capacitive scanner is used to measure the fuselage vibration response on a deHaviland Dash-8 fuselage, due to an internal noise source. The approach is validated by comparing this reciprocal noise transmission measurement with the direct measurement. The fuselage noise transmission information is then combined with computer predictions of the propeller pressure field data to predict the internal noise at two points.

Maritime patrol aircraft engine study, General Electric Derivative Engines. Volume III. Appendix B: performance data, TF 34/T7 study AL turboprop. Final report October 1978-April 1979

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hirschkron, R.; Davis, R.H.; Warren, R.E.

1979-04-30

This study developed data on General Electric common core derivative engines for use in Maritime Patrol Aircraft (MPA) concept formulation studies. The study included the screening of potential General Electric turbofan and turboprop/turboshaft engines and the preparation of technical and planning information on three of the most promising engine candidates. Screening of General Electric derivative candidates was performed utilizing an analytical MPA model using synthesized mission profiles to rank the candidates in terms of fuel consumption, weight, cost and complexity. The three turboprop engines selected for further study were as follows: TF34 growth derivative version with boost and new LPTmore » (TF34/T7 Study A1), F404 derivative with booster stages and new LPT (F404/T1 Study A1), and GE27 scaled and boosted study engine (GE27/T3 Study A1). Volume I summarizes the screening analysis and contains technical, planning, installation, cost and development data for the three selected turboprop engines. Volumes II, III and IV of this report contain the detailed performance data estimates for the GE27/T3 Study A1, TF34/T7 Study A1 and F404/T1 Study A1 turboprop engines, respectively.« less

Structure of ATP-Bound Human ATP:Cobalamin Adenosyltransferase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schubert,H.; Hill, C.

Mutations in the gene encoding human ATP:cobalamin adenosyltransferase (hATR) can result in the metabolic disorder known as methylmalonic aciduria (MMA). This enzyme catalyzes the final step in the conversion of cyanocobalamin (vitamin B{sub 12}) to the essential human cofactor adenosylcobalamin. Here we present the 2.5 {angstrom} crystal structure of ATP bound to hATR refined to an R{sub free} value of 25.2%. The enzyme forms a tightly associated trimer, where the monomer comprises a five-helix bundle and the active sites lie on the subunit interfaces. Only two of the three active sites within the trimer contain the bound ATP substrate, therebymore » providing examples of apo- and substrate-bound-active sites within the same crystal structure. Comparison of the empty and occupied sites indicates that twenty residues at the enzyme's N-terminus become ordered upon binding of ATP to form a novel ATP-binding site and an extended cleft that likely binds cobalamin. The structure explains the role of 20 invariant residues; six are involved in ATP binding, including Arg190, which hydrogen bonds to ATP atoms on both sides of the scissile bond. Ten of the hydrogen bonds are required for structural stability, and four are in positions to interact with cobalamin. The structure also reveals how the point mutations that cause MMA are deficient in these functions.« less

Maritime patrol aircraft engine study, General Electric Derivative Engines. Volume II. Appendix A: performance data, Ge27/T3 study A1 turboprop. Final report, October 1978-April 1979

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hirschkron, R.; Davis, R.H.; Warren, R.E.

1979-04-30

This study developed data on General Electric common core derivative engines for use in Maritime Patrol Aircraft (MPA) concept formulation studies. The study included the screening of potential General Electric turbofan and turboprop/turboshaft engines and the preparation of technical and planning information on three of the most promising engine candidates. Screening of General Electric derivative candidates was performed utilizing an analytical MPA model using synthesized mission profiles to rank the candidates in terms of fuel consumption, weight, cost and complexity. The three turboprop engines selected for further study were as follows: TF34 growth derivative version with boost and new LPTmore » (TF34/T7 Study A1), F404 derivative with booster stages and new LPT (F404/T1 Study A1), and GE27 scaled and boosted study engine (GE27/T3 Study A1). Volume I summarizes the screening analysis and contains technical, planning, installation, cost and development data for the three selected turboprop engines. Volumes II, III and IV of this report contain the detailed performance data estimates for the GE27/T3 Study A1, TF34/T7 Study A1 and F404/T1 Study A1 turboprop engines, respectively.« less

Maritime patrol aircraft engine study, General Electric Derivative Engines. Volume IV. Apendix C: performance data, F404/T1 study A1 turboprop. Final report, October 1978-April 1979

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hirschkron, R.; Davis, R.H.; Warren, R.E.

1979-04-30

This study developed data on General Electric common core derivative engines for use in Maritime Patrol Aircraft (MPA) concept formulation studies. The study included the screening of potential General Electric turbofan and turboprop/turboshaft engines and the preparation of technical and planning information on three of the most promising engine candidates. Screening of General Electric derivative candidates was performed utilizing an analytical MPA model using synthesized mission profiles to rank the candidates in terms of fuel consumption, weight, cost and complexity. The three turboprop engines selected for further study were as follows: TF34 growth derivative version with boost and new LPTmore » (TF34/T7 Study A1), F404 derivative with booster stages and new LPT (F404/T1 Study A1), and GE27 scaled and boosted study engine (GE27/T3 Study A1). Volume I summarizes the screening analysis and contains technical, planning, installation, cost and development data for the three selected turboprop engines. Volumes II, III and IV of this report contain the detailed performance data estimates for the GE27/T3 Study A1, TF34/T7 Study A1 and F404/T1 Study A1 turboprop engines, respectively.« less

Fuel conservation merits of advanced turboprop transport aircraft

NASA Technical Reports Server (NTRS)

Revell, J. D.; Tullis, R. H.

1977-01-01

The advantages of a propfan powered aircraft for the commercial air transportation system were assessed by the comparison with an equivalent turbofan transport. Comparisons were accomplished on the basis of fuel utilization and operating costs, as well as aircraft weight and size. Advantages of the propfan aircraft, concerning fuel utilization and operating costs, were accomplished by considering: (1) incorporation of propfan performance and acoustic data; (2) revised mission profiles (longer design range and reduction in; and cruise speed) (3) utilization of alternate and advanced technology engines.

Mutations in the Atp1p and Atp3p subunits of yeast ATP synthase differentially affect respiration and fermentation in Saccharomyces cerevisiae.

PubMed

Francis, Brian R; White, Karen H; Thorsness, Peter E

2007-04-01

ATP1-111, a suppressor of the slow-growth phenotype of yme1Delta lacking mitochondrial DNA is due to the substitution of phenylalanine for valine at position 111 of the alpha-subunit of mitochondrial ATP synthase (Atp1p in yeast). The suppressing activity of ATP1-111 requires intact beta (Atp2p) and gamma (Atp3p) subunits of mitochondrial ATP synthase, but not the stator stalk subunits b (Atp4p) and OSCP (Atp5p). ATP1-111 and other similarly suppressing mutations in ATP1 and ATP3 increase the growth rate of wild-type strains lacking mitochondrial DNA. These suppressing mutations decrease the growth rate of yeast containing an intact mitochondrial chromosome on media requiring oxidative phosphorylation, but not when grown on fermentable media. Measurement of chronological aging of yeast in culture reveals that ATP1 and ATP3 suppressor alleles in strains that contain mitochondrial DNA are longer lived than the isogenic wild-type strain. In contrast, the chronological life span of yeast cells lacking mitochondrial DNA and containing these mutations is shorter than that of the isogenic wild-type strain. Spore viability of strains bearing ATP1-111 is reduced compared to wild type, although ATP1-111 enhances the survival of spores that lacked mitochondrial DNA.

Flight investigation of cabin noise control treatments for a light turboprop aircraft

NASA Technical Reports Server (NTRS)

Wilby, J. F.; Oneal, R. L.; Mixson, J. S.

1985-01-01

The in-flight evaluation of noise control treatments for a light, twin-engined turboprop aircraft presents several problems associated with data analysis and interpretation. These problems include data repeatability, propeller synchronization, spatial distributions of the exterior pressure field and acoustic treatment, and the presence of flanking paths. They are discussed here with regard to a specific aeroplane configuration. Measurements were made in an untreated cabin and in a cabin fitted with an experimental sidewall treatment. Results are presented in terms of the insertion loss provided by the treatment and comparison made with predictions based on laboratory measurements.

ATP7A-related copper transport diseases-emerging concepts and future trends.

PubMed

Kaler, Stephen G

2011-01-01

This Review summarizes recent advances in understanding copper-transporting ATPase 1 (ATP7A), and examines the neurological phenotypes associated with dysfunction of this protein. Involvement of ATP7A in axonal outgrowth, synapse integrity and neuronal activation underscores the fundamental importance of copper metabolism to neurological function. Defects in ATP7A cause Menkes disease, an infantile-onset, lethal condition. Neonatal diagnosis and early treatment with copper injections enhance survival in patients with this disease, and can normalize clinical outcomes if mutant ATP7A molecules retain small amounts of residual activity. Gene replacement rescues a mouse model of Menkes disease, suggesting a potential therapeutic approach for patients with complete loss-of-function ATP7A mutations. Remarkably, a newly discovered ATP7A disorder-isolated distal motor neuropathy-has none of the characteristic clinical or biochemical abnormalities of Menkes disease or its milder allelic variant occipital horn syndrome (OHS), instead resembling Charcot-Marie-Tooth disease type 2. These findings indicate that ATP7A has a crucial but previously unappreciated role in motor neuron maintenance, and that the mechanism underlying ATP7A-related distal motor neuropathy is distinct from Menkes disease and OHS pathophysiology. Collectively, these insights refine our knowledge of the neurology of ATP7A-related copper transport diseases and pave the way for further progress in understanding ATP7A function.

Fatigue life analysis of a turboprop reduction gearbox

NASA Technical Reports Server (NTRS)

Lewicki, D. G.; Black, J. D.; Savage, M.; Coy, J. J.

1985-01-01

A fatigue life analysis of the Allison T56/501 turboprop reduction gearbox was developed. The life and reliability of the gearbox was based on the lives and reliabilities of the main power train bearings and gears. The bearing and gear lives were determined using the Lundberg-Palmgren theory and a mission profile. The five planet bearing set had the shortest calculated life among the various gearbox components, which agreed with field experience where the planet bearing had the greatest incidences of failure. The analytical predictions of relative lives among the various bearings were in reasonable agreement with field experience. The predicted gearbox life was in excellent agreement with field data when the material life adjustment factors alone were used. The gearbox had a lower predicted life in comparison with field data when no life adjustment factors were used or when lubrication life adjustment factors were used either alone or in combination with the material factors.

Fatigue life analysis of a turboprop reduction gearbox

NASA Technical Reports Server (NTRS)

Lewicki, D. G.; Coy, J. J.; Black, J. D.; Savage, M.

1986-01-01

A fatigue life analysis of the Allison T56/501 turboprop reduction gearbox was developed. The life and reliability of the gearbox was based on the lives and reliabilities of the main power train bearings and gears. The bearing and gear lives were determined using the Lundberg-Palmgren theory and a mission profile. The five planet bearing set had the shortest calculated life among the various gearbox components, which agreed with field experience where the planet bearing had the greatest incidences of failure. The analytical predictions of relative lives among the various bearings were in reasonable agreement with field experience. The predicted gearbox life was in excellent agreement with field data when the material life adjustment factors alone were used. The gearbox had a lower predicted life in comparison with field data when no life adjustment factors were used or when lubrication life adjustment factors were used either alone or in combination with the material factors.

Numerical and flight measured interior noise characteristics of a twin-engine turboprop general aviation aircraft

NASA Astrophysics Data System (ADS)

Marulo, F.; Lecce, L.; de Rosa, S.; D'Amato, C. A.; Verde, G.

The paper presents the flight test results of an interior noise measurement campaign on a twin-engine turboprop general aviation aircraft conducted for assessing the real values inside such aircraft and for approaching the problem of its noise reduction. Simultaneously a numerical study has been performed in order to correlate the experimental and the theoretical values, trying to come out with some guidelines for possible improvements without increasing excessively the costs of such study.

Structureborne noise control in advanced turboprop aircraft

NASA Technical Reports Server (NTRS)

Loeffler, Irvin J.

1987-01-01

Structureborne noise is discussed as a contributor to propeller aircraft interior noise levels that are nonresponsive to the application of a generous amount of cabin sidewall acoustic treatment. High structureborne noise levels may jeopardize passenger acceptance of the fuel-efficient high-speed propeller transport aircraft designed for cruise at Mach 0.65 to 0.85. These single-rotation tractor and counter-rotation tractor and pusher propulsion systems will consume 15 to 30 percent less fuel than advanced turbofan systems. Structureborne noise detection methodologies and the importance of development of a structureborne noise sensor are discussed. A structureborne noise generation mechanism is described in which the periodic components or propeller swirl produce periodic torques and forces on downstream wings and airfoils that are propagated to the cabin interior as noise. Three concepts for controlling structureborne noise are presented: (1) a stator row swirl remover, (2) selection of a proper combination of blade numbers in the rotor/stator system of a single-rotation propeller, and the rotor/rotor system of a counter-rotation propeller, and (3) a tuned mechanical absorber.

Future Propulsion Opportunities for Commuter Airplanes

NASA Technical Reports Server (NTRS)

Strack, W. C.

1982-01-01

Commuter airplane propulsion opportunities are summarized. Consideration is given to advanced technology conventional turboprop engines, advanced propellers, and several unconventional alternatives: regenerative turboprops, rotaries, and diesels. Advanced versions of conventional turboprops (including propellers) offer 15-20 percent savings in fuel and 10-15 percent in DOC compared to the new crop of 1500-2000 SHP engines currently in development. Unconventional engines could boost the fuel savings to 30-40 percent. The conclusion is that several important opportunities exist and, therefore, powerplant technology need not plateau.

Role of the P-Type ATPases, ATP7A and ATP7B in brain copper homeostasis.

PubMed

Telianidis, Jonathon; Hung, Ya Hui; Materia, Stephanie; Fontaine, Sharon La

2013-01-01

Over the past two decades there have been significant advances in our understanding of copper homeostasis and the pathological consequences of copper dysregulation. Cumulative evidence is revealing a complex regulatory network of proteins and pathways that maintain copper homeostasis. The recognition of copper dysregulation as a key pathological feature in prominent neurodegenerative disorders such as Alzheimer's, Parkinson's, and prion diseases has led to increased research focus on the mechanisms controlling copper homeostasis in the brain. The copper-transporting P-type ATPases (copper-ATPases), ATP7A and ATP7B, are critical components of the copper regulatory network. Our understanding of the biochemistry and cell biology of these complex proteins has grown significantly since their discovery in 1993. They are large polytopic transmembrane proteins with six copper-binding motifs within the cytoplasmic N-terminal domain, eight transmembrane domains, and highly conserved catalytic domains. These proteins catalyze ATP-dependent copper transport across cell membranes for the metallation of many essential cuproenzymes, as well as for the removal of excess cellular copper to prevent copper toxicity. A key functional aspect of these copper transporters is their copper-responsive trafficking between the trans-Golgi network and the cell periphery. ATP7A- and ATP7B-deficiency, due to genetic mutation, underlie the inherited copper transport disorders, Menkes and Wilson diseases, respectively. Their importance in maintaining brain copper homeostasis is underscored by the severe neuropathological deficits in these disorders. Herein we will review and update our current knowledge of these copper transporters in the brain and the central nervous system, their distribution and regulation, their role in normal brain copper homeostasis, and how their absence or dysfunction contributes to disturbances in copper homeostasis and neurodegeneration.

Role of the P-Type ATPases, ATP7A and ATP7B in brain copper homeostasis

PubMed Central

Telianidis, Jonathon; Hung, Ya Hui; Materia, Stephanie; Fontaine, Sharon La

2013-01-01

Over the past two decades there have been significant advances in our understanding of copper homeostasis and the pathological consequences of copper dysregulation. Cumulative evidence is revealing a complex regulatory network of proteins and pathways that maintain copper homeostasis. The recognition of copper dysregulation as a key pathological feature in prominent neurodegenerative disorders such as Alzheimer’s, Parkinson’s, and prion diseases has led to increased research focus on the mechanisms controlling copper homeostasis in the brain. The copper-transporting P-type ATPases (copper-ATPases), ATP7A and ATP7B, are critical components of the copper regulatory network. Our understanding of the biochemistry and cell biology of these complex proteins has grown significantly since their discovery in 1993. They are large polytopic transmembrane proteins with six copper-binding motifs within the cytoplasmic N-terminal domain, eight transmembrane domains, and highly conserved catalytic domains. These proteins catalyze ATP-dependent copper transport across cell membranes for the metallation of many essential cuproenzymes, as well as for the removal of excess cellular copper to prevent copper toxicity. A key functional aspect of these copper transporters is their copper-responsive trafficking between the trans-Golgi network and the cell periphery. ATP7A- and ATP7B-deficiency, due to genetic mutation, underlie the inherited copper transport disorders, Menkes and Wilson diseases, respectively. Their importance in maintaining brain copper homeostasis is underscored by the severe neuropathological deficits in these disorders. Herein we will review and update our current knowledge of these copper transporters in the brain and the central nervous system, their distribution and regulation, their role in normal brain copper homeostasis, and how their absence or dysfunction contributes to disturbances in copper homeostasis and neurodegeneration. PMID:23986700

Atomic model for the dimeric FO region of mitochondrial ATP synthase.

PubMed

Guo, Hui; Bueler, Stephanie A; Rubinstein, John L

2017-11-17

Mitochondrial adenosine triphosphate (ATP) synthase produces the majority of ATP in eukaryotic cells, and its dimerization is necessary to create the inner membrane folds, or cristae, characteristic of mitochondria. Proton translocation through the membrane-embedded F O region turns the rotor that drives ATP synthesis in the soluble F 1 region. Although crystal structures of the F 1 region have illustrated how this rotation leads to ATP synthesis, understanding how proton translocation produces the rotation has been impeded by the lack of an experimental atomic model for the F O region. Using cryo-electron microscopy, we determined the structure of the dimeric F O complex from Saccharomyces cerevisiae at a resolution of 3.6 angstroms. The structure clarifies how the protons travel through the complex, how the complex dimerizes, and how the dimers bend the membrane to produce cristae. Copyright © 2017, American Association for the Advancement of Science.

Measurements of propeller noise in a light turboprop airplane

NASA Technical Reports Server (NTRS)

Wilby, J. F.; Wilby, E. G.

1987-01-01

In-flight acoustic measurements have been made on the exterior and interior of a twin-engined turboprop airplane under controlled conditions to study data repeatability. It is found that the variability of the harmonic sound pressure levels in the cabin is greater than that for the exterior sound pressure levels, typical values for the standard deviation being +2.0 dB and -4.2 dB for the interior, versus +1.4 dB and -2.3 dB for the exterior. When insertion losses are determined for acoustic treatments in the cabin, the standard deviations of the data are typically + or - 6.5 dB. It is concluded that additional factors, such as accurate and repeatable selection of relative phase between propellers, controlled cabin-air-temperatures, installation of baseline acoustic absorption, and measurement of aircraft attitude, should be considered in order to reduce uncertainty in the measured data.

The outlook for advanced transport aircraft

NASA Technical Reports Server (NTRS)

Leavens, J. M., Jr.; Schaufele, R. D.; Jones, R. T.; Steiner, J. E.; Beteille, R.; Titcomb, G. A.; Coplin, J. F.; Rowe, B. H.; Lloyd-Jones, D. J.; Overend, W. J.

1982-01-01

The technological advances most likely to contribute to advanced aircraft designs and the efficiency, performance, and financial considerations driving the development directions for new aircraft are reviewed. Fuel-efficiency is perceived as the most critical factor for any new aircraft or component design, with most gains expected to come in areas of propulsion, aerodynamics, configurations, structural designs and materials, active controls, digital avionics, laminar flow control, and air-traffic control improvements. Any component area offers an efficiency improvement of 3-12%, with a maximum of 50% possible with a 4000 m range aircraft. Advanced turboprops have potential applications in short and medium haul subsonic aircraft, while a fuel efficient SST may be possible by the year 2000. Further discussion is devoted to the pivoted oblique wing aircraft, lightweight structures, and the necessity for short payback times.

Treatment of heterotopic ossification through remote ATP hydrolysis.

PubMed

Peterson, Jonathan R; De La Rosa, Sara; Eboda, Oluwatobi; Cilwa, Katherine E; Agarwal, Shailesh; Buchman, Steven R; Cederna, Paul S; Xi, Chuanwu; Morris, Michael D; Herndon, David N; Xiao, Wenzhong; Tompkins, Ronald G; Krebsbach, Paul H; Wang, Stewart C; Levi, Benjamin

2014-09-24

Heterotopic ossification (HO) is the pathologic development of ectopic bone in soft tissues because of a local or systemic inflammatory insult, such as burn injury or trauma. In HO, mesenchymal stem cells (MSCs) are inappropriately activated to undergo osteogenic differentiation. Through the correlation of in vitro assays and in vivo studies (dorsal scald burn with Achilles tenotomy), we have shown that burn injury enhances the osteogenic potential of MSCs and causes ectopic endochondral heterotopic bone formation and functional contractures through bone morphogenetic protein-mediated canonical SMAD signaling. We further demonstrated a prevention strategy for HO through adenosine triphosphate (ATP) hydrolysis at the burn site using apyrase. Burn site apyrase treatment decreased ATP, increased adenosine 3',5'-monophosphate, and decreased phosphorylation of SMAD1/5/8 in MSCs in vitro. This ATP hydrolysis also decreased HO formation and mitigated functional impairment in vivo. Similarly, selective inhibition of SMAD1/5/8 phosphorylation with LDN-193189 decreased HO formation and increased range of motion at the injury site in our burn model in vivo. Our results suggest that burn injury-exacerbated HO formation can be treated through therapeutics that target burn site ATP hydrolysis and modulation of SMAD1/5/8 phosphorylation. Copyright © 2014, American Association for the Advancement of Science.

Mechanisms of charge transfer in human copper ATPases ATP7A and ATP7B.

PubMed

Tadini-Buoninsegni, Francesco; Smeazzetto, Serena

2017-04-01

ATP7A and ATP7B are Cu + -transporting ATPases of subclass IB and play a fundamental role in intracellular copper homeostasis. ATP7A/B transfer Cu + ions across the membrane from delivery to acceptor proteins without establishing a free Cu + gradient. Transfer of copper across the membrane is coupled to ATP hydrolysis. Current measurements on solid supported membranes (SSM) were performed to investigate the mechanism of copper-related charge transfer across ATP7A and ATP7B. SSM measurements demonstrated that electrogenic copper displacement occurs within ATP7A/B following addition of ATP and formation of the phosphorylated intermediate. Comparison of the time constants for cation displacement in ATP7A/B and sarcoplasmic reticulum Ca 2+ -ATPase is consistent with the slower phosphoenzyme formation in copper ATPases. Moreover, ATP-dependent copper transfer in ATP7A/B is not affected by varying the pH, suggesting that net proton counter-transport may not occur in copper ATPases. Platinum anticancer drugs activate ATP7A/B and are subjected to ATP-dependent vectorial displacement with a mechanism analogous to that of copper. © 2016 IUBMB Life, 69(4):218-225, 2017. © 2017 International Union of Biochemistry and Molecular Biology.

Evaluation of advanced lift concepts and potential fuel conservation for short-haul aircraft

NASA Technical Reports Server (NTRS)

Sweet, H. S.; Renshaw, J. H.; Bowden, M. K.

1975-01-01

The effect of different field lengths, cruise requirements, noise level, and engine cycle characteristics on minimizing fuel consumption and minimizing operating cost at high fuel prices were evaluated for some advanced short-haul aircraft. The conceptual aircraft were designed for 148 passengers using the upper surface-internally blown jet flap, the augmentor wing, and the mechanical flap lift systems. Advanced conceptual STOL engines were evaluated as well as a near-term turbofan and turboprop engine. Emphasis was given to designs meeting noise levels equivalent to 95-100 EPNdB at 152 m (500 ft) sideline.

ATP synthase.

PubMed

Junge, Wolfgang; Nelson, Nathan

2015-01-01

Oxygenic photosynthesis is the principal converter of sunlight into chemical energy. Cyanobacteria and plants provide aerobic life with oxygen, food, fuel, fibers, and platform chemicals. Four multisubunit membrane proteins are involved: photosystem I (PSI), photosystem II (PSII), cytochrome b6f (cyt b6f), and ATP synthase (FOF1). ATP synthase is likewise a key enzyme of cell respiration. Over three billion years, the basic machinery of oxygenic photosynthesis and respiration has been perfected to minimize wasteful reactions. The proton-driven ATP synthase is embedded in a proton tight-coupling membrane. It is composed of two rotary motors/generators, FO and F1, which do not slip against each other. The proton-driven FO and the ATP-synthesizing F1 are coupled via elastic torque transmission. Elastic transmission decouples the two motors in kinetic detail but keeps them perfectly coupled in thermodynamic equilibrium and (time-averaged) under steady turnover. Elastic transmission enables operation with different gear ratios in different organisms.

Contraction of small mesenteric arteries induced by micromolar concentrations of ATP released from caged ATP.

PubMed

Sjöblom-Widfeldt, N; Arner, A; Nilsson, H

1993-01-01

The concentration dependence of ATP-induced contractions in isolated resistance arteries was estimated using photolysis of caged ATP. Rat mesenteric vessels were isolated and mounted for force registration in a small chamber allowing illumination from a xenon-flash lamp. Photolysis of 100 microM caged ATP, which released about 20 microM ATP within a few milliseconds in the vessel, induced a transient contraction with an amplitude approximately 40-50% of the response induced by 10 microM noradrenaline. The responses could neither be induced by the light flash as such nor by caged ATP alone nor by photolysis of caged phosphate. The amplitude of the contractions was dependent on the concentration of caged ATP, and the effective concentration for ATP was estimated to be in the range of 1-10 microM. In contrast, when ATP was introduced by diffusion, about a 100-fold higher concentration was required. Thus photolytic release of ATP minimizes metabolism before its action on receptors and reveals action of ATP in a concentration range consistent with a role of ATP as a transmitter in nervous regulation of the tone of resistance vessels.

Inhibition of plasma membrane Ca(2+)-ATPase by CrATP. LaATP but not CrATP stabilizes the Ca(2+)-occluded state.

PubMed

Moreira, Otacilio C; Rios, Priscila F; Barrabin, Hector

2005-07-15

The bidentate complex of ATP with Cr(3+), CrATP, is a nucleotide analog that is known to inhibit the sarcoplasmic reticulum Ca(2+)-ATPase and the Na(+),K(+)-ATPase, so that these enzymes accumulate in a conformation with the transported ion (Ca(2+) and Na(+), respectively) occluded from the medium. Here, it is shown that CrATP is also an effective and irreversible inhibitor of the plasma membrane Ca(2+)-ATPase. The complex inhibited with similar efficiency the Ca(2+)-dependent ATPase and the phosphatase activities as well as the enzyme phosphorylation by ATP. The inhibition proceeded slowly (T(1/2)=30 min at 37 degrees C) with a K(i)=28+/-9 microM. The inclusion of ATP, ADP or AMPPNP in the inhibition medium effectively protected the enzyme against the inhibition, whereas ITP, which is not a PMCA substrate, did not. The rate of inhibition was strongly dependent on the presence of Mg(2+) but unaltered when Ca(2+) was replaced by EGTA. In spite of the similarities with the inhibition of other P-ATPases, no apparent Ca(2+) occlusion was detected concurrent with the inhibition by CrATP. In contrast, inhibition by the complex of La(3+) with ATP, LaATP, induced the accumulation of phosphoenzyme with a simultaneous occlusion of Ca(2+) at a ratio close to 1.5 mol/mol of phosphoenzyme. The results suggest that the transport of Ca(2+) promoted by the plasma membrane Ca(2+)-ATPase goes through an enzymatic phospho-intermediate that maintains Ca(2+) ions occluded from the media. This intermediate is stabilized by LaATP but not by CrATP.

Imaging Adenosine Triphosphate (ATP)

PubMed Central

Rajendran, Megha; Dane, Eric; Conley, Jason; Tantama, Mathew

2016-01-01

Adenosine triphosphate (ATP) is a universal mediator of metabolism and signaling across unicellular and multicellular species. There is a fundamental interdependence between the dynamics of ATP and the physiology that occurs inside and outside the cell. Characterizing and understanding ATP dynamics provides valuable mechanistic insight into processes that range from neurotransmission to the chemotaxis of immune cells. Therefore, we require the methodology to interrogate both temporal and spatial components of ATP dynamics from the subcellular to organismal levels in live specimens. Over the last several decades, a number of molecular probes that are specific for ATP have been developed. These probes have been combined with imaging approaches, particularly optical microscopy, to enable qualitative and quantitative detection of this critical molecule. In this review, we survey current examples of technologies that are available to visualize ATP in living cells and identify areas where new tools and approaches are needed to expand our capabilities. PMID:27638696

Imaging Adenosine Triphosphate (ATP).

PubMed

Rajendran, Megha; Dane, Eric; Conley, Jason; Tantama, Mathew

2016-08-01

Adenosine triphosphate (ATP) is a universal mediator of metabolism and signaling across unicellular and multicellular species. There is a fundamental interdependence between the dynamics of ATP and the physiology that occurs inside and outside the cell. Characterizing and understanding ATP dynamics provide valuable mechanistic insight into processes that range from neurotransmission to the chemotaxis of immune cells. Therefore, we require the methodology to interrogate both temporal and spatial components of ATP dynamics from the subcellular to the organismal levels in live specimens. Over the last several decades, a number of molecular probes that are specific to ATP have been developed. These probes have been combined with imaging approaches, particularly optical microscopy, to enable qualitative and quantitative detection of this critical molecule. In this review, we survey current examples of technologies available for visualizing ATP in living cells, and identify areas where new tools and approaches are needed to expand our capabilities. © 2016 Marine Biological Laboratory.

Conformational dynamics of ATP/Mg:ATP in motor proteins via data mining and molecular simulation.

PubMed

Bojovschi, A; Liu, Ming S; Sadus, Richard J

2012-08-21

The conformational diversity of ATP/Mg:ATP in motor proteins was investigated using molecular dynamics and data mining. Adenosine triphosphate (ATP) conformations were found to be constrained mostly by inter cavity motifs in the motor proteins. It is demonstrated that ATP favors extended conformations in the tight pockets of motor proteins such as F(1)-ATPase and actin whereas compact structures are favored in motor proteins such as RNA polymerase and DNA helicase. The incorporation of Mg(2+) leads to increased flexibility of ATP molecules. The differences in the conformational dynamics of ATP/Mg:ATP in various motor proteins was quantified by the radius of gyration. The relationship between the simulation results and those obtained by data mining of motor proteins available in the protein data bank is analyzed. The data mining analysis of motor proteins supports the conformational diversity of the phosphate group of ATP obtained computationally.

Propeller installation effects on turboprop aircraft acoustics

NASA Astrophysics Data System (ADS)

Chirico, Giulia; Barakos, George N.; Bown, Nicholas

2018-06-01

Propeller installation options for a twin-engined turboprop aircraft are evaluated at cruise conditions, aiming to identify the quieter configuration. Computational fluid dynamics is used to investigate the near-field acoustics and transfer functions are employed to estimate the interior cabin noise. Co-rotating and counter-rotating installation options are compared. The effect of propeller synchrophasing is also considered. The employed method captures the complexity of the acoustic field generated by the interactions of the propeller sound fields among each other and with the airframe, showing also the importance of simulating the whole problem to predict the actual noise on a flying aircraft. Marked differences among the various layouts are observed. The counter-rotating top-in option appears the best in terms of acoustics, the top-out propeller rotation leading to louder noise because of inflow conditions and the occurrence of constructive acoustic interferences. Synchrophasing is shown to be beneficial for co-rotating propellers, specially regarding the interior noise, because of favorable effects in the interaction between the propeller direct sound field and the noise due to the airframe. An angle closer to the maximum relative blade shift was found to be the best choice, yielding, however, higher sound levels than those provided by the counter-rotating top-in layout.

Noise abatement technology options for conventional turboprop airplanes. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Galloway, W.J.; Wilby, J.F.

1981-06-01

The practical application of noise control technology to new and derivative conventional turboprop airplanes likely to come into service in the 1980's has been analyzed with a view to determining noise control cost/benefits. The analysis identifies feasible noise control methods, applies them to four study airplanes, and presents the noise reductions in terms of the equivalent perceived noise level at takeoff, sideline and approach locations, and the effect on the area within selected EPNL contours. Noise reductions of up to 8.3 dB for takeoff and 10.7 dB for approach are calculated for the study airplanes but, for most cases, themore » changes are less than 5 dB. Weight and cost increases associated with the noise control treatments are determined under the assumption there they are no changes to airplane performance or fuel consumption.« less

Low-speed wind-tunnel tests of an advanced eight-bladed propeller

NASA Technical Reports Server (NTRS)

Coe, P. L., Jr.; Gentry, G. L., Jr.; Dunham, D. M.

1985-01-01

As part of a research program on advanced turboprop aircraft aerodynamics, a low-speed wind-tunnel investigation was conducted to document the basic performance and force and moment characteristics of an advanced eight-bladed propeller. The results show that in addition to the normal force and pitching moment produced by the propeller/nacelle combination at angle of attack, a significant side force and yawing moment are also produced. Furthermore, it is shown that for test conditions wherein compressibility effects can be ignored, accurate simulation of propeller performance and flow fields can be achieved by matching the nondimensional power loading of the model propeller to that of the full-scale propeller.

Caged ATP - an internal calibration method for ATP bioluminescence assays.

PubMed

Calvert, R M; Hopkins, H C; Reilly, M J; Forsythe, S J

2000-03-01

ATP bioluminescence, based on the firefly luciferase system, is used for the rapid determination of hygienic practices in the food industry. This study has demonstrated the use of caged ATP as an internal ATP standard and quantified the effects of industrial cleansing solutions, alcoholic beverages and pH on firefly luciferase activity. The light signal was quenched 6-47% by five cleansing solutions at standard working concentrations. Ethanol at 1% (v/v) inhibited bioluminescence by 15% (w/v) whereas concentrations above 4% enhanced the light output. The light signal was quenched by 20-25% at pH values below pH 4 and above pH 10.

Unified aeroacoustics analysis for high speed turboprop aerodynamics and noise. Volume 2: Development of theory for wing shielding

NASA Technical Reports Server (NTRS)

Amiet, R. K.

1991-01-01

A unified theory for aerodynamics and noise of advanced turboprops is presented. The theory and a computer code developed for evaluation at the shielding benefits that might be expected by an aircraft wing in a wing-mounted propeller installation are presented. Several computed directivity patterns are presented to demonstrate the theory. Recently with the advent of the concept of using the wing of an aircraft for noise shielding, the case of diffraction by a surface in a flow has been given attention. The present analysis is based on the case of diffraction of no flow. By combining a Galilean and a Lorentz transform, the wave equation with a mean flow can be reduced to the ordinary equation. Allowance is also made in the analysis for the case of a swept wing. The same combination of Galilean and Lorentz transforms lead to a problem with no flow but a different sweep. The solution procedures for the cases of leading and trailing edges are basically the same. Two normalizations of the solution are given by the computer program. FORTRAN computer programs are presented with detailed documentation. The output from these programs compares favorably with the results of other investigators.

Installed nacelle drag-improvement tests of an M = 0.8 turboprop transport configuration

NASA Technical Reports Server (NTRS)

Levin, A. D.; Smith, R. C.

1983-01-01

An unpowered semispan model of a representative turboprop configuration was tested to determine the effect of configuration modifications on the the nonmetric body and wing juncture. It is indicated that the jet off nacelle-installation drag can be approximately 25% of the cruise drag. However, the losses can be reduced to 17% by changes to the wing leading edge and nacelle intersection. Comparison of test results from a semispan nonmetric fuselage model with those from a full span metric fuselage show differences in angles of attack produced the same lift. It is found that the constant lift drag rise of the semispan model is higher because of the increased angle of attack to achieve the same lift.

Noise transmission through sidewall treatments applicable to twin-engine turboprop aircraft

NASA Technical Reports Server (NTRS)

Grosveld, F. W.

1983-01-01

The noise transmission loss characteristics of the sidewall treatment in the propeller plane of a twin-engine turboprop aircraft are experimentally investigated in the NASA Langley Research Center Transmission Loss Facility. The sound attenuation properties of the individual elements of this treatment are evaluated showing least noise transmission loss in the low frequencies (below 500 Hz) where the excitation levels at the propeller blade passage frequency and the first few harmonics are highest. It is shown that single and double wall resonances play an important role in the noise transmission loss values of the treatment at these low frequencies suggesting that a limp mass with a very low resonance frequency serves better as a trim panel than a trim panel having a high structural stiffness. It is indicated that the window structures might be a potential noise control problem.

Noise transmission through sidewall treatments applicable to twin-engine turboprop aircraft

NASA Astrophysics Data System (ADS)

Grosveld, F. W.

1983-04-01

The noise transmission loss characteristics of the sidewall treatment in the propeller plane of a twin-engine turboprop aircraft are experimentally investigated in the NASA Langley Research Center Transmission Loss Facility. The sound attenuation properties of the individual elements of this treatment are evaluated showing least noise transmission loss in the low frequencies (below 500 Hz) where the excitation levels at the propeller blade passage frequency and the first few harmonics are highest. It is shown that single and double wall resonances play an important role in the noise transmission loss values of the treatment at these low frequencies suggesting that a limp mass with a very low resonance frequency serves better as a trim panel than a trim panel having a high structural stiffness. It is indicated that the window structures might be a potential noise control problem.

15 CFR 295.11 - Technical and educational services for ATP recipients.

Code of Federal Regulations, 2010 CFR

2010-01-01

... Foreign Trade NATIONAL INSTITUTE OF STANDARDS AND TECHNOLOGY, DEPARTMENT OF COMMERCE NIST EXTRAMURAL PROGRAMS ADVANCED TECHNOLOGY PROGRAM General § 295.11 Technical and educational services for ATP recipients... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Technical and educational services for...

The ATP/DNA Ratio Is a Better Indicator of Islet Cell Viability Than the ADP/ATP Ratio

PubMed Central

Suszynski, T.M.; Wildey, G.M.; Falde, E.J.; Cline, G.W.; Maynard, K. Stewart; Ko, N.; Sotiris, J.; Naji, A.; Hering, B.J.; Papas, K.K.

2009-01-01

Real-time, accurate assessment of islet viability is critical for avoiding transplantation of nontherapeutic preparations. Measurements of the intracellular ADP/ATP ratio have been recently proposed as useful prospective estimates of islet cell viability and potency. However, dead cells may be rapidly depleted of both ATP and ADP, which would render the ratio incapable of accounting for dead cells. Since the DNA of dead cells is expected to remain stable over prolonged periods of time (days), we hypothesized that use of the ATP/DNA ratio would take into account dead cells and may be a better indicator of islet cell viability than the ADP/ATP ratio. We tested this hypothesis using mixtures of healthy and lethally heat-treated (HT) rat insulinoma cells and human islets. Measurements of ATP/DNA and ADP/ATP from the known mixtures of healthy and HT cells and islets were used to evaluate how well these parameters correlated with viability. The results indicated that ATP and ADP were rapidly (within 1 hour) depleted in HT cells. The fraction of HT cells in a mixture correlated linearly with the ATP/DNA ratio, whereas the ADP/ADP ratio was highly scattered, remaining effectively unchanged. Despite similar limitations in both ADP/ADP and ATP/DNA ratios, in that ATP levels may fluctuate significantly and reversibly with metabolic stress, the results indicated that ATP/DNA was a better measure of islet viability than the ADP/ATP ratio. PMID:18374063

Large-scale Advanced Prop-fan (LAP) technology assessment report

NASA Technical Reports Server (NTRS)

Degeorge, C. L.

1988-01-01

The technologically significant findings and accomplishments of the Large Scale Advanced Prop-Fan (LAP) program in the areas of aerodynamics, aeroelasticity, acoustics and materials and fabrication are described. The extent to which the program goals related to these disciplines were achieved is discussed, and recommendations for additional research are presented. The LAP program consisted of the design, manufacture and testing of a near full-scale Prop-Fan or advanced turboprop capable of operating efficiently at speeds to Mach .8. An aeroelastically scaled model of the LAP was also designed and fabricated. The goal of the program was to acquire data on Prop-Fan performance that would indicate the technology readiness of Prop-Fans for practical applications in commercial and military aviation.

Contribution of extracellular ATP on the cell-surface F1F0-ATP synthase-mediated intracellular triacylglycerol accumulation.

PubMed

Kita, Toshiyuki; Arakaki, Naokatu

2015-01-01

Cell-surface F1F0-ATP synthase was involved in the cell signaling mediating various biological functions. Recently, we found that cell-surface F1F0-ATP synthase plays a role on intracellular triacylglycerol accumulation in adipocytes, and yet, the underlying mechanisms remained largely unknown. In this study, we investigated the role of extracellular ATP on the intracellular triacylglycerol accumulation. We demonstrated that significant amounts of ATP were produced extracellularly by cultured 3T3-L1 adipocytes and that the antibodies against α and β subunits of F1F0-ATP synthase inhibited the extracellular ATP production. Piceatannol, a F1F0-ATP synthase inhibitor, and apyrase, an enzyme which degrades extracellular ATP, suppressed triacylglycerol accumulation. The selective P2Y1 receptor antagonist MRS2500 significantly inhibited triacylglycerol accumulation, whereas the selective P2X receptor antagonist NF279 has less effect. The present results indicate that cell-surface F1F0-ATP synthase on adipocytes is functional in extracellular ATP production and that the extracellular ATP produced contributes, at least in part, to the cell-surface F1F0-ATP synthase-mediated intracellular triacylglycerol accumulation in adipocytes through P2Y1 receptor.

Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging

PubMed Central

Yaginuma, Hideyuki; Kawai, Shinnosuke; Tabata, Kazuhito V.; Tomiyama, Keisuke; Kakizuka, Akira; Komatsuzaki, Tamiki; Noji, Hiroyuki; Imamura, Hiromi

2014-01-01

Recent advances in quantitative single-cell analysis revealed large diversity in gene expression levels between individual cells, which could affect the physiology and/or fate of each cell. In contrast, for most metabolites, the concentrations were only measureable as ensemble averages of many cells. In living cells, adenosine triphosphate (ATP) is a critically important metabolite that powers many intracellular reactions. Quantitative measurement of the absolute ATP concentration in individual cells has not been achieved because of the lack of reliable methods. In this study, we developed a new genetically-encoded ratiometric fluorescent ATP indicator “QUEEN”, which is composed of a single circularly-permuted fluorescent protein and a bacterial ATP binding protein. Unlike previous FRET-based indicators, QUEEN was apparently insensitive to bacteria growth rate changes. Importantly, intracellular ATP concentrations of numbers of bacterial cells calculated from QUEEN fluorescence were almost equal to those from firefly luciferase assay. Thus, QUEEN is suitable for quantifying the absolute ATP concentration inside bacteria cells. Finally, we found that, even for a genetically-identical Escherichia coli cell population, absolute concentrations of intracellular ATP were significantly diverse between individual cells from the same culture, by imaging QUEEN signals from single cells. PMID:25283467

Can advanced technology improve future commuter aircraft

NASA Technical Reports Server (NTRS)

Williams, L. J.; Snow, D. B.

1981-01-01

The short-haul service abandoned by the trunk and local airlines is being picked up by the commuter airlines using small turboprop-powered aircraft. Most of the existing small transport aircraft currently available represent a relatively old technology level. However, several manufacturers have initiated the development of new or improved commuter transport aircraft. These aircraft are relatively conservative in terms of technology. An examination is conducted of advanced technology to identify those technologies that, if developed, would provide the largest improvements for future generations of these aircraft. Attention is given to commuter aircraft operating cost, aerodynamics, structures and materials, propulsion, aircraft systems, and technology integration. It is found that advanced technology can improve future commuter aircraft and that the largest of these improvements will come from the synergistic combination of technological advances in all of the aircraft disciplines. The most important goals are related to improved fuel efficiency and increased aircraft productivity.

ATP analogues at a glance.

PubMed

Bagshaw, C

2001-02-01

ATP has long been known to play a central role in the energetics of cells both in transduction mechanisms and in metabolic pathways, and is involved in regulation of enzyme, channel and receptor activities. Numerous ATP analogues have been synthesised to probe the role of ATP in biosystems (Yount, 1975; Jameson and Eccleston, 1997; Bagshaw, 1998). In general, two contrasting strategies are employed. Modifications may be introduced deliberately to change the properties of ATP (e.g. making it non-hydrolysable) so as to perturb the chemical steps involved in its action. Typically these involve modification of the phosphate chain. Alternatively, derivatives (e.g. fluorescent probes) are designed to report on the action of ATP but have a minimal effect on its properties. ATP-utilising systems vary enormously in their specificity; so what acts as a good analogue in one case may be very poor in another. The accompanying poster shows a representative selection of derivatives that have been synthesised and summarises their key properties.

Optogenetic control of ATP release

NASA Astrophysics Data System (ADS)

Lewis, Matthew A.; Joshi, Bipin; Gu, Ling; Feranchak, Andrew; Mohanty, Samarendra K.

2013-03-01

Controlled release of ATP can be used for understanding extracellular purinergic signaling. While coarse mechanical forces and hypotonic stimulation have been utilized in the past to initiate ATP release from cells, these methods are neither spatially accurate nor temporally precise. Further, these methods cannot be utilized in a highly effective cell-specific manner. To mitigate the uncertainties regarding cellular-specificity and spatio-temporal release of ATP, we herein demonstrate use of optogenetics for ATP release. ATP release in response to optogenetic stimulation was monitored by Luciferin-Luciferase assay (North American firefly, photinus pyralis) using luminometer as well as mesoscopic bioluminescence imaging. Our result demonstrates repetitive release of ATP subsequent to optogenetic stimulation. It is thus feasible that purinergic signaling can be directly detected via imaging if the stimulus can be confined to single cell or in a spatially-defined group of cells. This study opens up new avenue to interrogate the mechanisms of purinergic signaling.

Erythrocyte haemolysate interacts with ATP-Fe to form a complex containing iron, ATP and 13 800 MW polypeptide.

PubMed

Weaver, J; Zhan, H; Pollack, S

1993-01-01

Iron first entering the reticulocyte is bound to ATP in the low MW cytosolic pool; some is also 'loosely bound' to haemoglobin, coeluting with haemoglobin from a molecular sieve column though not incorporated into haem. When haemolysate is mixed with ATP-Fe in vitro a similar high MW iron-containing complex is formed: the ATP-Fe interacts with a non-haemoglobin constituent of the haemolysate to form a high MW ATP-Fe complex in which the ratio of ATP:Fe (originally 6:1) is reversed, so that the complex contains more iron than ATP. The high MW ATP-Fe complex is formed even when ATP is in 150-fold molar excess and is formed without detectable hydrolysis of the ATP. The activity of haemolysate in forming the high MW ATP-Fe complex is not diminished by dialysis; all of the activity is recovered in the haemoglobin-containing fraction obtained from an Ultrogel AcA 44 column. The activity does not derive from haemoglobin since 85% of the activity is removed when haemoglobin is purified from haemolysate with DEAE-Sephadex. The chelatable iron pool of the cell probably includes both the high MW ATP-Fe complex and low MW ATP-Fe. Shunting of ATP-Fe to a high MW aggregate reduces the amount of iron present in the highly reactive low MW form and thus probably serves to limit the formation of cell damaging radicals.

ATP Maintenance via Two Types of ATP Regulators Mitigates Pathological Phenotypes in Mouse Models of Parkinson's Disease.

PubMed

Nakano, Masaki; Imamura, Hiromi; Sasaoka, Norio; Yamamoto, Masamichi; Uemura, Norihito; Shudo, Toshiyuki; Fuchigami, Tomohiro; Takahashi, Ryosuke; Kakizuka, Akira

2017-08-01

Parkinson's disease is assumed to be caused by mitochondrial dysfunction in the affected dopaminergic neurons in the brain. We have recently created small chemicals, KUSs (Kyoto University Substances), which can reduce cellular ATP consumption. By contrast, agonistic ligands of ERRs (estrogen receptor-related receptors) are expected to raise cellular ATP levels via enhancing ATP production. Here, we show that esculetin functions as an ERR agonist, and its addition to culture media enhances glycolysis and mitochondrial respiration, leading to elevated cellular ATP levels. Subsequently, we show the neuroprotective efficacies of KUSs, esculetin, and GSK4716 (an ERRγ agonist) against cell death in Parkinson's disease models. In the surviving neurons, ATP levels and expression levels of α-synuclein and CHOP (an ER stress-mediated cell death executor) were all rectified. We propose that maintenance of ATP levels, by inhibiting ATP consumption or enhancing ATP production, or both, would be a promising therapeutic strategy for Parkinson's disease. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

ATP Dependence of Na+/H+ Exchange

PubMed Central

Demaurex, Nicolas; Romanek, Robert R.; Orlowski, John; Grinstein, Sergio

1997-01-01

We studied the ATP dependence of NHE-1, the ubiquitous isoform of the Na+/H+ antiporter, using the whole-cell configuration of the patch-clamp technique to apply nucleotides intracellularly while measuring cytosolic pH (pHi) by microfluorimetry. Na+/H+ exchange activity was measured as the Na+-driven pHi recovery from an acid load, which was imposed via the patch pipette. In Chinese hamster ovary (CHO) fibroblasts stably transfected with NHE-1, omission of ATP from the pipette solution inhibited Na+/H+ exchange. Conversely, ATP perfusion restored exchange activity in cells that had been metabolically depleted by 2-deoxy-d-glucose and oligomycin. In cells dialyzed in the presence of ATP, no “run-down” was observed even after extended periods, suggesting that the nucleotide is the only diffusible factor required for optimal NHE-1 activity. Half-maximal activation of the antiporter was obtained at ∼5 mM Mg-ATP. Submillimolar concentrations failed to sustain Na+/H+ exchange even when an ATP regenerating system was included in the pipette solution. High ATP concentrations are also known to be required for the optimal function of other cation exchangers. In the case of the Na/Ca2+ exchanger, this requirement has been attributed to an aminophospholipid translocase, or “flippase.” The involvement of this enzyme in Na+/H+ exchange was examined using fluorescent phosphatidylserine, which is actively translocated by the flippase. ATP depletion decreased the transmembrane uptake of NBD-labeled phosphatidylserine (NBD-PS), indicating that the flippase was inhibited. Diamide, an agent reported to block the flippase, was as potent as ATP depletion in reducing NBD-PS uptake. However, diamide had no effect on Na+/H+ exchange, implying that the effect of ATP is not mediated by changes in lipid distribution across the plasma membrane. K-ATP and ATPγS were as efficient as Mg-ATP in sustaining NHE-1 activity, while AMP-PNP and AMP-PCP only partially substituted for ATP. In

Fuel conservative aircraft engine technology

NASA Technical Reports Server (NTRS)

Nored, D. L.

1978-01-01

Technology developments for more fuel-efficiency subsonic transport aircraft are reported. Three major propulsion projects were considered: (1) engine component improvement - directed at current engines; (2) energy efficient engine - directed at new turbofan engines; and (3) advanced turboprops - directed at technology for advanced turboprop-powered aircraft. Each project is reviewed and some of the technologies and recent accomplishments are described.

Clusterin and COMMD1 Independently Regulate Degradation of the Mammalian Copper ATPases ATP7A and ATP7B*

PubMed Central

Materia, Stephanie; Cater, Michael A.; Klomp, Leo W. J.; Mercer, Julian F. B.; La Fontaine, Sharon

2012-01-01

ATP7A and ATP7B are copper-transporting P1B-type ATPases (Cu-ATPases) that are critical for regulating intracellular copper homeostasis. Mutations in the genes encoding ATP7A and ATP7B lead to copper deficiency and copper toxicity disorders, Menkes and Wilson diseases, respectively. Clusterin and COMMD1 were previously identified as interacting partners of these Cu-ATPases. In this study, we confirmed that clusterin and COMMD1 interact to down-regulate both ATP7A and ATP7B. Overexpression and knockdown of clusterin/COMMD1 decreased and increased, respectively, endogenous levels of ATP7A and ATP7B, consistent with a role in facilitating Cu-ATPase degradation. We demonstrate that whereas the clusterin/ATP7B interaction was enhanced by oxidative stress or mutation of ATP7B, the COMMD1/ATP7B interaction did not change under oxidative stress conditions, and only increased with ATP7B mutations that led to its misfolding. Clusterin and COMMD1 facilitated the degradation of ATP7B containing the same Wilson disease-causing C-terminal mutations via different degradation pathways, clusterin via the lysosomal pathway and COMMD1 via the proteasomal pathway. Furthermore, endogenous ATP7B existed in a complex with clusterin and COMMD1, but these interactions were neither competitive nor cooperative and occurred independently of each other. Together these data indicate that clusterin and COMMD1 represent alternative and independent systems regulating Cu-ATPase quality control, and consequently contributing to the maintenance of copper homeostasis. PMID:22130675

In-flight acoustic measurements on a light twin-engined turboprop airplane

NASA Technical Reports Server (NTRS)

Wilby, J. F.; Mcdaniel, C. D.; Wilby, E. G.

1985-01-01

Four series of flight tests were conducted to measure sound pressure levels inside and outside the cabin of a twin-engined turboprop airplane. Particular emphasis was placed on harmonics of the propeller blade passage frequency. The cabin was unfurnished for the first three flights, when the main objective was to investigate the repeatability of the data. For the fourth flight, the cabin was treated with fiberglass batts. Typically, the exterior sound pressure levels were found to vary 3 to 5 dB for a given harmonic, but variations as high as 8 dB were observed. The variability of harmonic levels within the cabin was slightly higher but depended on control of the relative phase between the propellers; when phase was not controlled the average variability was about 10 dB. Noise reductions provided by the fuselage structure were in the range of 20 to 40 dB, when an exterior microphone in the plane of rotation of the propeller was used as reference.

Inhibition of ATP Synthase by Chlorinated Adenosine Analogue

PubMed Central

Chen, Lisa S.; Nowak, Billie J.; Ayres, Mary L.; Krett, Nancy L.; Rosen, Steven T.; Zhang, Shuxing; Gandhi, Varsha

2009-01-01

8-Chloroadenosine (8-Cl-Ado) is a ribonucleoside analogue that is currently in clinical trial for chronic lymphocytic leukemia. Based on the decline in cellular ATP pool following 8-Cl-Ado treatment, we hypothesized that 8-Cl-ADP and 8-Cl-ATP may interfere with ATP synthase, a key enzyme in ATP production. Mitochondrial ATP synthase is composed of two major parts; FO intermembrane base and F1 domain, containing α and β subunits. Crystal structures of both α and β subunits that bind to the substrate, ADP, are known in tight binding (αdpβdp) and loose binding (αtpβtp) states. Molecular docking demonstrated that 8-Cl-ADP/8-Cl-ATP occupied similar binding modes as ADP/ATP in the tight and loose binding sites of ATP synthase, respectively, suggesting that the chlorinated nucleotide metabolites may be functional substrates and inhibitors of the enzyme. The computational predictions were consistent with our whole cell biochemical results. Oligomycin, an established pharmacological inhibitor of ATP synthase, decreased both ATP and 8-Cl-ATP formation from exogenous substrates, however, did not affect pyrimidine nucleoside analogue triphosphate accumulation. Synthesis of ATP from ADP was inhibited in cells loaded with 8-Cl-ATP. These biochemical studies are in consent with the computational modeling; in the αtpβtp state 8-Cl-ATP occupies similar binding as ANP, a non-hydrolyzable ATP mimic that is a known inhibitor. Similarly, in the substrate binding site (αdpβdp) 8-Cl-ATP occupies a similar position as ATP mimic ADP-BeF3 −. Collectively, our current work suggests that 8-Cl-ADP may serve as a substrate and the 8-Cl-ATP may be an inhibitor of ATP synthase. PMID:19477165

An experimental evaluation of S-duct inlet-diffuser configurations for turboprop offset gearbox applications

NASA Technical Reports Server (NTRS)

Mcdill, Paul L.

1986-01-01

A test program, utilizing a large scale model, was run in the NASA Lewis Research Center 10- by 10-ft wind tunnel to examine the influence on performance of design parameters of turboprop S-duct inlet/diffuser systems. The parametric test program investigated inlet lip thickness, inlet/diffuser cross-sectional geometry, throat design Mach number, and shaft fairing shape. The test program was run at angles of attack to 15 deg and tunnel Mach numbers to 0.35. Results of the program indicate that current design techniques can be used to design inlet/diffuser systems with acceptable total pressure recovery, but several of the design parameters, notably lip thickness (contraction ratio) and shaft fairing cross section, must be optimized to prevent excessive distortion at the compressor face.

Acoustic flight testing of advanced design propellers on a JetStar aircraft

NASA Technical Reports Server (NTRS)

Lasagna, P.; Mackall, K.

1981-01-01

Advanced turboprop-powered aircraft have the potential to reduce fuel consumption by 15 to 30 percent as compared with an equivalent technology turbofan-powered aircraft. An important obstacle to the use of advanced design propellers is the cabin noise generated at Mach numbers up to .8 and at altitudes up to 35,000 feet. As part of the NASA Aircraft Energy Efficiency Program, the near-field acoustic characteristics on a series of advanced design propellers are investigated. Currently, Dryden Flight Research Center is flight testing a series of propellers on a JetStar airplane. The propellers used in the flight test were previously tested in wind tunnels at the Lewis Research Center. Data are presented showing the narrow band spectra, acoustic wave form, and acoustic contours on the fuselage surface. Additional flights with the SR-3 propeller and other advanced propellers are planned in the future.

[P4-ATP-ase Atp8b1/FIC1: structural properties and (patho)physiological functions].

PubMed

Korneenko, T V; Pestov, N B; Okkelman, I A; Modyanov, N N; Shakhparonov, M I

2015-01-01

P4-ATP-ases comprise an interesting family among P-type ATP-ases, since they are thought to play a major role in the transfer of phospholipids such as phosphatydylserine from the outer leaflet to the inner leaflet. Isoforms of P4-ATP-ases are partially interchangeable but peculiarities of tissue-specific expression of their genes, intracellular localization of proteins, as well as regulatory pathways lead to the fact that, on the organismal level, serious pathologies may develop in the presence of structural abnormalities in certain isoforms. Among P4-ATP-ases a special place is occupied by ATP8B1, for which several mutations are known that lead to serious hereditary diseases: two forms of congenital cholestasis (PFIC1 or Byler disease and benign recurrent intrahepatic cholestasis) with extraliver symptoms such as sensorineural hearing loss. The physiological function of the Atp8b1/FIC1 protein is known in general outline: it is responsible for transport of certain phospholipids (phosphatydylserine, cardiolipin) for the outer monolayer of the plasma membrane to the inner one. It is well known that perturbation of membrane asymmetry, caused by the lack of Atp8B1 activity, leads to death of hairy cells of the inner ear, dysfunction of bile acid transport in liver-cells that causes cirrhosis. It is also probable that insufficient activity of Atp8b1/FIC1 increases susceptibility to bacterial pneumonia.Regulatory pathways of Atp8b1/FIC1 activity in vivo remain to be insufficiently studied and this opens novel perspectives for research in this field that may allow better understanding of molecular processes behind the development of certain pathologies and to reveal novel therapeutical targets.

Variable effects of the mitoK(ATP) channel modulators diazoxide and 5-HD in ATP-depleted renal epithelial cells.

PubMed

Nilakantan, Vani; Liang, Huanling; Mortensen, Jordan; Taylor, Erin; Johnson, Christopher P

2010-02-01

The role of mitochondrial K(ATP) (mitoK(ATP)) channels in renal ischemia-reperfusion injury is controversial with studies showing both protective and deleterious effects. In this study, we compared the effects of the putative mitoK(ATP) opener, diazoxide, and the mitoK(ATP) blocker, 5-hydroxydecanoate (5-HD) on cytotoxicity and apoptosis in tubular epithelial cells derived from rat (NRK-52E) and pig (LLC-PK1) following in vitro ischemic injury. Following ATP depletion-recovery, there was a significant increase in cytotoxicity in both NRK cells and LLC-PK1 cells although NRK cells were more sensitive to the injury. Diazoxide treatment attenuated cytotoxicity in both cell types and 5-HD treatment-increased cytotoxicity in the sensitive NRK cells in a superoxide-dependant manner. The protective effect of diazoxide was also reversed in the presence of 5-HD in ATP-depleted NRK cells. The ATP depletion-mediated increase in superoxide was enhanced by both diazoxide and 5-HD with the effect being more pronounced in the cells undergoing 5-HD treatment. Further, ATP depletion-induced activation of caspase-3 was decreased by diazoxide in NRK cells. In order to determine the signaling pathways involved in apoptosis, we examined the activation of Erk and JNK in ATP-depleted NRK cells. Diazoxide-activated Erk in ATP-depleted cells, but did not have any effect on JNK activation. In contrast, 5-HD did not impact Erk levels but increased JNK activation even under controlled conditions. Further, the use of a JNK inhibitor with 5-HD reversed the deleterious effects of 5-HD. This study demonstrates that in cells that are sensitive to ATP depletion-recovery, mitoK(ATP) channels protect against ATP depletion-mediated cytotoxicity and apoptosis through Erk- and JNK-dependant mechanisms.

Sound propagation elements in evaluation of en route noise of advanced turbofan aircraft

NASA Technical Reports Server (NTRS)

Sutherland, Louis C.; Wesler, John

1990-01-01

Cruise noise from an advanced turboprop aircraft is reviewed on the basis of available wind tunnel data to estimate the aircraft noise signature at the source. Available analytical models are used to evaluate the sound levels at the ground. The analysis allows reasonable estimates to be made of the community noise levels that might be generated during cruise by such aircraft, provides the basis for preliminary comparisons with available data on noise of existing aircraft during climb and helps to identify the dominant elements of the sound propagation models applicable to this situation.

Oral Adenosine-5'-triphosphate (ATP) Administration Increases Postexercise ATP Levels, Muscle Excitability, and Athletic Performance Following a Repeated Sprint Bout.

PubMed

Purpura, Martin; Rathmacher, John A; Sharp, Matthew H; Lowery, Ryan P; Shields, Kevin A; Partl, Jeremy M; Wilson, Jacob M; Jäger, Ralf

2017-01-01

Oral adenosine-5'-triphosphate (ATP) administration has failed to increase plasma ATP levels; however, chronic supplementation with ATP has shown to increase power, strength, lean body mass, and blood flow in trained athletes. The purpose of this study was to investigate the effects of ATP supplementation on postexercise ATP levels and on muscle activation and excitability and power following a repeated sprint bout. In a double-blind, placebo-controlled, randomized design, 42 healthy male individuals were given either 400 mg of ATP as disodium salt or placebo for 2 weeks prior to an exercise bout. During the exercise bout, muscle activation and excitability (ME, ratio of power output to muscle activation) and Wingate test peak power were measured during all sprints. ATP and metabolites were measured at baseline, after supplementation, and immediately following exercise. Oral ATP supplementation prevented a drop in ATP, adenosine-5'-diphosphate (ADP), and adenosine-5'-monophosphate (AMP) levels postexercise (p < 0.05). No group by time interaction was observed for muscle activation. Following the supplementation period, muscle excitability significantly decreased in later bouts 8, 9, and 10 in the placebo group (-30.5, -28.3, and -27.9%, respectively; p < 0.02), whereas ATP supplementation prevented the decline in later bouts. ATP significantly increased Wingate peak power in later bouts compared to baseline (bout 8: +18.3%, bout 10: +16.3%). Oral ATP administration prevents exercise-induced declines in ATP and its metabolite and enhances peak power and muscular excitability, which may be beneficial for sports requiring repeated high-intensity sprinting bouts.

Mitochondrial flashes regulate ATP homeostasis in the heart

PubMed Central

Wang, Xianhua; Zhang, Xing; Wu, Di; Huang, Zhanglong; Hou, Tingting; Jian, Chongshu; Yu, Peng; Lu, Fujian; Zhang, Rufeng; Sun, Tao; Li, Jinghang; Qi, Wenfeng; Wang, Yanru; Gao, Feng; Cheng, Heping

2017-01-01

The maintenance of a constant ATP level (‘set-point’) is a vital homeostatic function shared by eukaryotic cells. In particular, mammalian myocardium exquisitely safeguards its ATP set-point despite 10-fold fluctuations in cardiac workload. However, the exact mechanisms underlying this regulation of ATP homeostasis remain elusive. Here we show mitochondrial flashes (mitoflashes), recently discovered dynamic activity of mitochondria, play an essential role for the auto-regulation of ATP set-point in the heart. Specifically, mitoflashes negatively regulate ATP production in isolated respiring mitochondria and, their activity waxes and wanes to counteract the ATP supply-demand imbalance caused by superfluous substrate and altered workload in cardiomyocytes. Moreover, manipulating mitoflash activity is sufficient to inversely shift the otherwise stable ATP set-point. Mechanistically, the Bcl-xL-regulated proton leakage through F1Fo-ATP synthase appears to mediate the coupling between mitoflash production and ATP set-point regulation. These findings indicate mitoflashes appear to constitute a digital auto-regulator for ATP homeostasis in the heart. DOI: http://dx.doi.org/10.7554/eLife.23908.001 PMID:28692422

Evidence for the Synthesis of ATP by an F0F1 ATP Synthase in Membrane Vesicles from Halorubrum Saccharovorum

NASA Technical Reports Server (NTRS)

Faguy, David; Lawson, Darion; Hochstein, Lawrence I.; Chang, Sherwood (Technical Monitor)

1996-01-01

Vesicles prepared in a buffer containing ADP, Mg(2+) and Pi synthesized ATP at an initial rate of 2 nmols/min/mg protein after acidification of the bulk medium (pH 8 (right arrow) 4). The intravesicular ATP concentration reached a steady state after about 30 seconds and slowly declined thereafter. ATP synthesis was inhibited by low concentrations of dicyclohexylcarbodiimide and m-chlorophenylhydrazone indicating that synthesis took place in response to the proton gradient. NEM and PCMS, which inhibit vacuolar ATPases and the vacuolar-like ATPases of extreme halophiles, did not affect ATP synthesis, and, in fact, produced higher steady state levels of ATP. This suggested that two ATPase activities were present, one which catalyzed ATP synthesis and one that caused its hydrolysis. Azide, a specific inhibitor of F0F1 ATP Synthases, inhibited halobacterial ATP synthesis. The distribution of acridine orange as imposed by a delta pH demonstrated that azide inhibition was not due to the collapse of the proton gradient due to azide acting as a protonophore. Such an effect was observed, but only at azide concentrations higher than those that inhibited ATP synthesis. These results confirm the earler observations with cells of H. saccharovorum and other extreme halophiles that ATP synthesis is inconsistent with the operation of a vacuolar-like ATPase. Therefore, the observation that a vacuolar-like enzyme is responsible for ATP synthesis (and which serves as the basis for imputing ATP synthesis to the vacuolar-like ATPases of the extreme halophiles, and the Archaea in general) should be taken with some degree of caution.

Biallelic Mutations in ATP5F1D , which Encodes a Subunit of ATP Synthase, Cause a Metabolic Disorder

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oláhová, Monika; Yoon, Wan Hee; Thompson, Kyle

ATP synthase, H + transporting, mitochondrial F1 complex, δ subunit (ATP5F1D; formerly ATP5D) is a subunit of mitochondrial ATP synthase and plays an important role in coupling proton translocation and ATP production. Here, we describe two individuals, each with homozygous missense variants in ATP5F1D, who presented with episodic lethargy, metabolic acidosis, 3-methylglutaconic aciduria, and hyperammonemia. Subject 1, homozygous for c.245C>T (p.Pro82Leu), presented with recurrent metabolic decompensation starting in the neonatal period, and subject 2, homozygous for c.317T>G (p.Val106Gly), presented with acute encephalopathy in childhood. Cultured skin fibroblasts from these individuals exhibited impaired assembly of F 1F O ATP synthase andmore » subsequent reduced complex V activity. Cells from subject 1 also exhibited a significant decrease in mitochondrial cristae. Knockdown of Drosophila ATPsynδ, the ATP5F1D homolog, in developing eyes and brains caused a near complete loss of the fly head, a phenotype that was fully rescued by wild-type human ATP5F1D. In contrast, expression of the ATP5F1D c.245C>T and c.317T>G variants rescued the head-size phenotype but recapitulated the eye and antennae defects seen in other genetic models of mitochondrial oxidative phosphorylation deficiency. Our data establish c.245C>T (p.Pro82Leu) and c.317T>G (p.Val106Gly) in ATP5F1D as pathogenic variants leading to a Mendelian mitochondrial disease featuring episodic metabolic decompensation.« less

Biallelic Mutations in ATP5F1D, which Encodes a Subunit of ATP Synthase, Cause a Metabolic Disorder.

PubMed

Oláhová, Monika; Yoon, Wan Hee; Thompson, Kyle; Jangam, Sharayu; Fernandez, Liliana; Davidson, Jean M; Kyle, Jennifer E; Grove, Megan E; Fisk, Dianna G; Kohler, Jennefer N; Holmes, Matthew; Dries, Annika M; Huang, Yong; Zhao, Chunli; Contrepois, Kévin; Zappala, Zachary; Frésard, Laure; Waggott, Daryl; Zink, Erika M; Kim, Young-Mo; Heyman, Heino M; Stratton, Kelly G; Webb-Robertson, Bobbie-Jo M; Snyder, Michael; Merker, Jason D; Montgomery, Stephen B; Fisher, Paul G; Feichtinger, René G; Mayr, Johannes A; Hall, Julie; Barbosa, Ines A; Simpson, Michael A; Deshpande, Charu; Waters, Katrina M; Koeller, David M; Metz, Thomas O; Morris, Andrew A; Schelley, Susan; Cowan, Tina; Friederich, Marisa W; McFarland, Robert; Van Hove, Johan L K; Enns, Gregory M; Yamamoto, Shinya; Ashley, Euan A; Wangler, Michael F; Taylor, Robert W; Bellen, Hugo J; Bernstein, Jonathan A; Wheeler, Matthew T

2018-03-01

ATP synthase, H + transporting, mitochondrial F1 complex, δ subunit (ATP5F1D; formerly ATP5D) is a subunit of mitochondrial ATP synthase and plays an important role in coupling proton translocation and ATP production. Here, we describe two individuals, each with homozygous missense variants in ATP5F1D, who presented with episodic lethargy, metabolic acidosis, 3-methylglutaconic aciduria, and hyperammonemia. Subject 1, homozygous for c.245C>T (p.Pro82Leu), presented with recurrent metabolic decompensation starting in the neonatal period, and subject 2, homozygous for c.317T>G (p.Val106Gly), presented with acute encephalopathy in childhood. Cultured skin fibroblasts from these individuals exhibited impaired assembly of F 1 F O ATP synthase and subsequent reduced complex V activity. Cells from subject 1 also exhibited a significant decrease in mitochondrial cristae. Knockdown of Drosophila ATPsynδ, the ATP5F1D homolog, in developing eyes and brains caused a near complete loss of the fly head, a phenotype that was fully rescued by wild-type human ATP5F1D. In contrast, expression of the ATP5F1D c.245C>T and c.317T>G variants rescued the head-size phenotype but recapitulated the eye and antennae defects seen in other genetic models of mitochondrial oxidative phosphorylation deficiency. Our data establish c.245C>T (p.Pro82Leu) and c.317T>G (p.Val106Gly) in ATP5F1D as pathogenic variants leading to a Mendelian mitochondrial disease featuring episodic metabolic decompensation. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Biallelic Mutations in ATP5F1D , which Encodes a Subunit of ATP Synthase, Cause a Metabolic Disorder

DOE PAGES

Oláhová, Monika; Yoon, Wan Hee; Thompson, Kyle; ...

2018-02-22

ATP synthase, H + transporting, mitochondrial F1 complex, δ subunit (ATP5F1D; formerly ATP5D) is a subunit of mitochondrial ATP synthase and plays an important role in coupling proton translocation and ATP production. Here, we describe two individuals, each with homozygous missense variants in ATP5F1D, who presented with episodic lethargy, metabolic acidosis, 3-methylglutaconic aciduria, and hyperammonemia. Subject 1, homozygous for c.245C>T (p.Pro82Leu), presented with recurrent metabolic decompensation starting in the neonatal period, and subject 2, homozygous for c.317T>G (p.Val106Gly), presented with acute encephalopathy in childhood. Cultured skin fibroblasts from these individuals exhibited impaired assembly of F 1F O ATP synthase andmore » subsequent reduced complex V activity. Cells from subject 1 also exhibited a significant decrease in mitochondrial cristae. Knockdown of Drosophila ATPsynδ, the ATP5F1D homolog, in developing eyes and brains caused a near complete loss of the fly head, a phenotype that was fully rescued by wild-type human ATP5F1D. In contrast, expression of the ATP5F1D c.245C>T and c.317T>G variants rescued the head-size phenotype but recapitulated the eye and antennae defects seen in other genetic models of mitochondrial oxidative phosphorylation deficiency. Our data establish c.245C>T (p.Pro82Leu) and c.317T>G (p.Val106Gly) in ATP5F1D as pathogenic variants leading to a Mendelian mitochondrial disease featuring episodic metabolic decompensation.« less

Development of GUI Type On-Line Condition Monitoring Program for a Turboprop Engine Using Labview

NASA Astrophysics Data System (ADS)

Kong, Changduk; Kim, Keonwoo

2011-12-01

Recently, an aero gas turbine health monitoring system has been developed for precaution and maintenance action against faults or performance degradations of the advanced propulsion system which occurs in severe environments such as high altitude, foreign object damage particles, hot and heavy rain and snowy atmospheric conditions. However to establish this health monitoring system, the online condition monitoring program is firstly required, and the program must monitor the engine performance trend through comparison between measured engine performance data and base performance results calculated by base engine performance model. This work aims to develop a GUI type on-line condition monitoring program for the PT6A-67 turboprop engine of a high altitude and long endurance operation UAV using LabVIEW. The base engine performance of the on-line condition monitoring program is simulated using component maps inversely generated from the limited performance deck data provided by engine manufacturer. The base engine performance simulation program is evaluated because analysis results by this program agree well with the performance deck data. The proposed on-line condition program can monitor the real engine performance as well as the trend through precise comparison between clean engine performance results calculated by the base performance simulation program and measured engine performance signals. In the development phase of this monitoring system, a signal generation module is proposed to evaluate the proposed online monitoring system. For user friendly purpose, all monitoring program are coded by LabVIEW, and monitoring examples are demonstrated using the proposed GUI type on-condition monitoring program.

ECN-15662

NASA Image and Video Library

1981-05-21

The Dryden C-140 JetStar during testing of advanced propfan designs. Dryden conducted flight research in 1981-1982 on several designs. The technology was developed under the direction of the Lewis Research Center (today the Glenn Research Center, Cleveland, OH) under the Advanced Turboprop Program. Under that program, Langley Research Center in Virginia oversaw work on accoustics and noise reduction. These efforts were intended to develop a high-speed and fuel-efficient turboprop system.

Advanced general aviation comparative engine/airframe integration study

NASA Technical Reports Server (NTRS)

Huggins, G. L.; Ellis, D. R.

1981-01-01

The NASA Advanced Aviation Comparative Engine/Airframe Integration Study was initiated to help determine which of four promising concepts for new general aviation engines for the 1990's should be considered for further research funding. The engine concepts included rotary, diesel, spark ignition, and turboprop powerplants; a conventional state-of-the-art piston engine was used as a baseline for the comparison. Computer simulations of the performance of single and twin engine pressurized aircraft designs were used to determine how the various characteristics of each engine interacted in the design process. Comparisons were made of how each engine performed relative to the others when integrated into an airframe and required to fly a transportation mission.

Volume-dependent ATP-conductive large-conductance anion channel as a pathway for swelling-induced ATP release.

PubMed

Sabirov, R Z; Dutta, A K; Okada, Y

2001-09-01

In mouse mammary C127i cells, during whole-cell clamp, osmotic cell swelling activated an anion channel current, when the phloretin-sensitive, volume-activated outwardly rectifying Cl(-) channel was eliminated. This current exhibited time-dependent inactivation at positive and negative voltages greater than around +/-25 mV. The whole-cell current was selective for anions and sensitive to Gd(3)+. In on-cell patches, single-channel events appeared with a lag period of approximately 15 min after a hypotonic challenge. Under isotonic conditions, cell-attached patches were silent, but patch excision led to activation of currents that consisted of multiple large-conductance unitary steps. The current displayed voltage- and time-dependent inactivation similar to that of whole-cell current. Voltage-dependent activation profile was bell-shaped with the maximum open probability at -20 to 0 mV. The channel in inside-out patches had the unitary conductance of approximately 400 pS, a linear current-voltage relationship, and anion selectivity. The outward (but not inward) single-channel conductance was suppressed by extracellular ATP with an IC(50) of 12.3 mM and an electric distance (delta) of 0.47, whereas the inward (but not outward) conductance was inhibited by intracellular ATP with an IC(50) of 12.9 mM and delta of 0.40. Despite the open channel block by ATP, the channel was ATP-conductive with P(ATP)/P(Cl) of 0.09. The single-channel activity was sensitive to Gd(3)+, SITS, and NPPB, but insensitive to phloretin, niflumic acid, and glibenclamide. The same pharmacological pattern was found in swelling-induced ATP release. Thus, it is concluded that the volume- and voltage-dependent ATP-conductive large-conductance anion channel serves as a conductive pathway for the swelling-induced ATP release in C127i cells.

The Structural Basis of ATP as an Allosteric Modulator

PubMed Central

Wang, Qi; Shen, Qiancheng; Li, Shuai; Nussinov, Ruth; Zhang, Jian

2014-01-01

Adenosine-5’-triphosphate (ATP) is generally regarded as a substrate for energy currency and protein modification. Recent findings uncovered the allosteric function of ATP in cellular signal transduction but little is understood about this critical behavior of ATP. Through extensive analysis of ATP in solution and proteins, we found that the free ATP can exist in the compact and extended conformations in solution, and the two different conformational characteristics may be responsible for ATP to exert distinct biological functions: ATP molecules adopt both compact and extended conformations in the allosteric binding sites but conserve extended conformations in the substrate binding sites. Nudged elastic band simulations unveiled the distinct dynamic processes of ATP binding to the corresponding allosteric and substrate binding sites of uridine monophosphate kinase, and suggested that in solution ATP preferentially binds to the substrate binding sites of proteins. When the ATP molecules occupy the allosteric binding sites, the allosteric trigger from ATP to fuel allosteric communication between allosteric and functional sites is stemmed mainly from the triphosphate part of ATP, with a small number from the adenine part of ATP. Taken together, our results provide overall understanding of ATP allosteric functions responsible for regulation in biological systems. PMID:25211773

Dual recognition unit strategy improves the specificity of the adenosine triphosphate (ATP) aptamer biosensor for cerebral ATP assay.

PubMed

Yu, Ping; He, Xiulan; Zhang, Li; Mao, Lanqun

2015-01-20

Adenosine triphosphate (ATP) aptamer has been widely used as a recognition unit for biosensor development; however, its relatively poor specificity toward ATP against adenosine-5'-diphosphate (ADP) and adenosine-5'-monophosphate (AMP) essentially limits the application of the biosensors in real systems, especially in the complex cerebral system. In this study, for the first time, we demonstrate a dual recognition unit strategy (DRUS) to construct a highly selective and sensitive ATP biosensor by combining the recognition ability of aptamer toward A nucleobase and of polyimidazolium toward phosphate. The biosensors are constructed by first confining the polyimidazolium onto a gold surface by surface-initiated atom transfer radical polymerization (SI-ATRP), and then the aptamer onto electrode surface by electrostatic self-assembly to form dual-recognition-unit-functionalized electrodes. The constructed biosensor based on DRUS not only shows an ultrahigh sensitivity toward ATP with a detection limit down to the subattomole level but also an ultrahigh selectivity toward ATP without interference from ADP and AMP. The constructed biosensor is used for selective and sensitive sensing of the extracellular ATP in the cerebral system by combining in vivo microdialysis and can be used as a promising neurotechnology to probing cerebral ATP concentration.

An advanced pitch change mechanism incorporating a hybrid traction drive

NASA Technical Reports Server (NTRS)

Steinetz, B. M.; Loewenthal, S. H.; Sargisson, D. F.; White, G.

1984-01-01

A design of a propeller pitch control mechanism is described that meets the demanding requirements of a high-power, advanced turboprop. In this application, blade twisting moment torque can be comparable to that of the main reduction gearbox output: precise pitch control, reliability and compactness are all at a premium. A key element in the design is a compact, high-ratio hybrid traction drive which offers low torque ripple and high torsional stiffness. The traction drive couples a high speed electric motor/alternator unit to a ball screw that actuates the blade control links. The technical merits of this arrangement and the performance characteristics of the traction drive are discussed.

Thermodynamics of proton transport coupled ATP synthesis.

PubMed

Turina, Paola; Petersen, Jan; Gräber, Peter

2016-06-01

The thermodynamic H(+)/ATP ratio of the H(+)-ATP synthase from chloroplasts was measured in proteoliposomes after energization of the membrane by an acid base transition (Turina et al. 2003 [13], 418-422). The method is discussed, and all published data obtained with this system are combined and analyzed as a single dataset. This meta-analysis led to the following results. 1) At equilibrium, the transmembrane ΔpH is energetically equivalent to the transmembrane electric potential difference. 2) The standard free energy for ATP synthesis (reference reaction) is ΔG°(ref)=33.8±1.3kJ/mol. 3) The thermodynamic H(+)/ATP ratio, as obtained from the shift of the ATP synthesis equilibrium induced by changing the transmembrane ΔpH (varying either pH(in) or pH(out)) is 4.0±0.1. The structural H(+)/ATP ratio, calculated from the ratio of proton binding sites on the c-subunit-ring in F(0) to the catalytic nucleotide binding sites on the β-subunits in F(1), is c/β=14/3=4.7. We infer that the energy of 0.7 protons per ATP that flow through the enzyme, but do not contribute to shifting the ATP/(ADP·Pi) ratio, is used for additional processes within the enzyme, such as activation, and/or energy dissipation, due e.g. to internal uncoupling. The ratio between the thermodynamic and the structural H(+)/ATP values is 0.85, and we conclude that this value represents the efficiency of the chemiosmotic energy conversion within the chloroplast H(+)-ATP synthase. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorescent ATP analog mant-ATP reports dynein activity in the isolated Chlamydomonas axoneme

NASA Astrophysics Data System (ADS)

Feofilova, Maria; Howard, Jonathon

Eukaryotic flagella are long rod-like extensions of cells, which play a fundamental role in single cell movement, as well as in fluid transport. Flagella contain a highly evolutionary conserved mechanical structure called the axoneme. The motion of the flagellum is generated by dynein motor proteins located all along the length of the axoneme. How the force production of motors is controlled spatially and temporally is still an open question. Therefore, monitoring dynein activity in the axonemal structure is expected to provide novel insights in regulation of the beat. We use high sensitivity fluorescence microscopy to monitor the binding and hydrolysis kinetics of the fluorescently labeled ATP analogue mant-ATP (2'(3')-O-(N-methylanthraniloyl) adenosine 5'-triphosphate), which is known to support dynein activity. By studying the kinetics of mant-ATP fluorescence, we identified distinct mant-ATP binding sites in the axoneme. The application of this method to axonemes with reduced amounts of dynein, showed evidence that one of the sites is associated with binding to dynein. In the future, we would like to use this method to find the spatial distribution of dynein activity in the axoneme.

Performance and Weight Estimates for an Advanced Open Rotor Engine

NASA Technical Reports Server (NTRS)

Hendricks, Eric S.; Tong, Michael T.

2012-01-01

NASA s Environmentally Responsible Aviation Project and Subsonic Fixed Wing Project are focused on developing concepts and technologies which may enable dramatic reductions to the environmental impact of future generation subsonic aircraft. The open rotor concept (also historically referred to an unducted fan or advanced turboprop) may allow for the achievement of this objective by reducing engine fuel consumption. To evaluate the potential impact of open rotor engines, cycle modeling and engine weight estimation capabilities have been developed. The initial development of the cycle modeling capabilities in the Numerical Propulsion System Simulation (NPSS) tool was presented in a previous paper. Following that initial development, further advancements have been made to the cycle modeling and weight estimation capabilities for open rotor engines and are presented in this paper. The developed modeling capabilities are used to predict the performance of an advanced open rotor concept using modern counter-rotating propeller designs. Finally, performance and weight estimates for this engine are presented and compared to results from a previous NASA study of advanced geared and direct-drive turbofans.

Magnetic field affects enzymatic ATP synthesis.

PubMed

Buchachenko, Anatoly L; Kuznetsov, Dmitry A

2008-10-01

The rate of ATP synthesis by creatine kinase extracted from V. xanthia venom was shown to depend on the magnetic field. The yield of ATP produced by enzymes with 24Mg2+ and 26Mg2+ ions in catalytic sites increases by 7-8% at 55 mT and then decreases at 80 mT. For enzyme with 25Mg2+ ion in a catalytic site, the ATP yield increases by 50% and 70% in the fields 55 and 80 mT, respectively. In the Earth field the rate of ATP synthesis by enzyme, in which Mg2+ ion has magnetic nucleus 25Mg, is 2.5 times higher than that by enzymes, in which Mg2+ ion has nonmagnetic, spinless nuclei 24Mg or 26Mg. Both magnetic field effect and magnetic isotope effect demonstrate that the ATP synthesis is an ion-radical process, affected by Zeeman interaction and hyperfine coupling in the intermediate ion-radical pair.

Cervical anterior transpedicular screw fixation (ATPS)—Part II. Accuracy of manual insertion and pull-out strength of ATPS

PubMed Central

Acosta, Frank; Tauber, Mark; Fox, Michael; Martin, Hudelmaier; Forstner, Rosmarie; Augat, Peter; Penzkofer, Rainer; Pirich, Christian; Kässmann, H.; Resch, Herbert; Hitzl, Wolfgang

2008-01-01

Reconstruction after multilevel decompression of the cervical spine, especially in the weakened osteoporotic, neoplastic or infectious spine often requires circumferential stabilization and fusion. To avoid the additional posterior surgery in these cases while increasing rigidity of anterior-only screw-plate constructs, the authors introduce the concept of anterior transpedicular screw (ATPS) fixation. We demonstrated its morphological feasibility as well as its indications in a previous study in Part I of our project. Consequently, the objectives of the current study were to assess the ex vivo accuracy of placing ATPS into the cervical vertebra as well as the biomechanical performance of ATPS in comparison to traditional vertebral body screws (VBS) in terms of pull-out strength (POS). Twenty-three ATPS were inserted alternately to two screws into the pedicles and vertebral bodies, respectively, of six cadaveric specimens from C3–T1. For insertion of ATPS, a manual fluoroscopically assisted technique was used. Pre- and post insertional CT-scans were used to assess accuracy of ATPS insertion in the axial and sagittal planes. A newly designed grading system and accuracy score were used to delineate accuracy of ATPS insertion. Following insertion of screws, 23 ATPS and 22 VBS were subjected to pull-out testing (POT). The bone mineral density (BMD) of each specimen was assessed prior to POT. Statistical analysis showed that the incidence of correctly placed screws and non-critical pedicles breaches in axial plane was 78.3%, and 95.7% in sagittal plane. Hence, according to our definition of “critical” pedicle breach that exposes neurovascular structures at risk, 21.7% (n = 5) of all ATPS inserted showed a critical pedicle breach in axial plane. Notably, no critical pedicle perforation occurred at the C6 to T1 levels. Pull-out testing of ATPS and VBS revealed that pull-out resistance of ATPS was 2.5-fold that of VBS. Mean POS of 23 ATPS with a mean BMD of 0.566�

An ATP synthase harboring an atypical γ-subunit is involved in ATP synthesis in tomato fruit chromoplasts.

PubMed

Pateraki, Irini; Renato, Marta; Azcón-Bieto, Joaquín; Boronat, Albert

2013-04-01

Chromoplasts are non-photosynthetic plastids specialized in the synthesis and accumulation of carotenoids. During fruit ripening, chloroplasts differentiate into photosynthetically inactive chromoplasts in a process characterized by the degradation of the thylakoid membranes, and by the active synthesis and accumulation of carotenoids. This transition renders chromoplasts unable to photochemically synthesize ATP, and therefore these organelles need to obtain the ATP required for anabolic processes through alternative sources. It is widely accepted that the ATP used for biosynthetic processes in non-photosynthetic plastids is imported from the cytosol or is obtained through glycolysis. In this work, however, we show that isolated tomato (Solanum lycopersicum) fruit chromoplasts are able to synthesize ATP de novo through a respiratory pathway using NADPH as an electron donor. We also report the involvement of a plastidial ATP synthase harboring an atypical γ-subunit induced during ripening, which lacks the regulatory dithiol domain present in plant and algae chloroplast γ-subunits. Silencing of this atypical γ-subunit during fruit ripening impairs the capacity of isolated chromoplast to synthesize ATP de novo. We propose that the replacement of the γ-subunit present in tomato leaf and green fruit chloroplasts by the atypical γ-subunit lacking the dithiol domain during fruit ripening reflects evolutionary changes, which allow the operation of chromoplast ATP synthase under the particular physiological conditions found in this organelle. © 2013 The Authors The Plant Journal © 2013 Blackwell Publishing Ltd.

A taste for ATP: neurotransmission in taste buds

PubMed Central

Kinnamon, Sue C.; Finger, Thomas E.

2013-01-01

Not only is ATP a ubiquitous source of energy but it is also used widely as an intercellular signal. For example, keratinocytes release ATP in response to numerous external stimuli including pressure, heat, and chemical insult. The released ATP activates purinergic receptors on nerve fibers to generate nociceptive signals. The importance of an ATP signal in epithelial-to-neuronal signaling is nowhere more evident than in the taste system. The receptor cells of taste buds release ATP in response to appropriate stimulation by tastants and the released ATP then activates P2X2 and P2X3 receptors on the taste nerves. Genetic ablation of the relevant P2X receptors leaves an animal without the ability to taste any primary taste quality. Of interest is that release of ATP by taste receptor cells occurs in a non-vesicular fashion, apparently via gated membrane channels. Further, in keeping with the crucial role of ATP as a neurotransmitter in this system, a subset of taste cells expresses a specific ectoATPase, NTPDase2, necessary to clear extracellular ATP which otherwise will desensitize the P2X receptors on the taste nerves. The unique utilization of ATP as a key neurotransmitter in the taste system may reflect the epithelial rather than neuronal origins of the receptor cells. PMID:24385952

Dynamics of the metal binding domains and regulation of the human copper transporters ATP7B and ATP7A.

PubMed

Yu, Corey H; Dolgova, Natalia V; Dmitriev, Oleg Y

2017-04-01

Copper transporters ATP7A and ATP7B regulate copper levels in the human cells and deliver copper to the biosynthetic pathways. ATP7A and ATP7B belong to the P-type ATPases and share much of the domain architecture and the mechanism of ATP hydrolysis with the other, well-studied, enzymes of this type. A unique structural feature of the copper ATPases is the chain of six cytosolic metal-binding domains (MBDs), which are believed to be involved in copper-dependent regulation of the activity and intracellular localization of these enzymes. Although the structures of all the MBDs have been solved, the mechanism of copper-dependent regulation of ATP7B and ATP7A, the roles of individual MBDs, and the relationship between the regulatory and catalytic copper binding are still unknown. We describe the structure and dynamics of the MBDs, review the current knowledge about their functional roles and propose a mechanism of regulation of ATP7B by copper-dependent changes in the dynamics and conformation of the MBD chain. Transient interactions between the MBDs, rather than transitions between distinct static conformations are likely to form the structural basis of regulation of the ATP-dependent copper transporters in human cells. © 2016 IUBMB Life, 69(4):226-235, 2017. © 2017 International Union of Biochemistry and Molecular Biology.

ATP Synthesis in the Extremely Halophilic Bacteria

NASA Technical Reports Server (NTRS)

Hochstein, Lawrence I.; Morrison, David (Technical Monitor)

1994-01-01

The proton-translocating ATPases are multimeric enzymes that carry out a multitude of essential functions. Their origin and evolution represent a seminal event in the early evolution of life. Amino acid sequences of the two largest subunits from archaeal ATPases (A-ATPases), vacuolar ATPases (V-ATPases), and FOF1-ATP syntheses (FATPases) suggest these ATPases evolved from an ancestral vacuolar-like ATP syntheses. A necessary consequence of this notion is that the A-ATPases are ATP syntheses. With the possible exception of the A-ATPase from Halobacterium salinarium. no A-ATPase has been demonstrated to synthesize ATP. The evidence for this case is dubious since ATP synthesis occurs only when conditions are distinctively unphysiological. We demonstrated that ATP synthesis in H.saccharovorum is inconsistent with the operation of an A-type ATPase. In order to determine if this phenomenon was unique to H. saccharovorum, ATP synthesis was examined in various extremely halophilic bacteria with the goal of ascertaining if it resembled what occurred in a. saccharovorum, or was consistent with the operation of an A-type ATPase. A-, V-, and F-type ATPases respond singularly to certain inhibitors. Therefore, the effect of these inhibitors on ATP synthesis in several extreme halophiles was determined. Inhibitors that either blocked or collapsed proton-gradients inhibited the steady state synthesis of ATP thus verifying that synthesis took place at the expense of a proton gradient. Azide, an inhibitor of F-ATPases inhibited ATP synthesis. Since the arginine-dependent synthesis of ATP, which occurs by way of substrate-level phosphorylation, was unaffected by azide, it was unlikely that azide acted as an "uncoupler." N -ethylmaleimide and nitrate, which inhibit V- and A-ATPases, either did not inhibit ATP synthesis or resulted in higher steady-state levels of ATP. These results suggest there are two types of proton-motive ATPases in the extreme halophiles (and presumably in other

Snapshots of the maltose transporter during ATP hydrolysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oldham, Michael L.; Chen, Jue

2011-12-05

ATP-binding cassette transporters are powered by ATP, but the mechanism by which these transporters hydrolyze ATP is unclear. In this study, four crystal structures of the full-length wild-type maltose transporter, stabilized by adenosine 5{prime}-({beta},{gamma}-imido)triphosphate or ADP in conjunction with phosphate analogs BeF{sub 3}{sup -}, VO{sub 4}{sup 3-}, or AlF{sub 4}{sup -}, were determined to 2.2- to 2.4-{angstrom} resolution. These structures led to the assignment of two enzymatic states during ATP hydrolysis and demonstrate specific functional roles of highly conserved residues in the nucleotide-binding domain, suggesting that ATP-binding cassette transporters catalyze ATP hydrolysis via a general base mechanism.

Aeroacoustics of advanced propellers

NASA Technical Reports Server (NTRS)

Groeneweg, John F.

1990-01-01

The aeroacoustics of advanced, high speed propellers (propfans) are reviewed from the perspective of NASA research conducted in support of the Advanced Turboprop Program. Aerodynamic and acoustic components of prediction methods for near and far field noise are summarized for both single and counterrotation propellers in uninstalled and configurations. Experimental results from tests at both takeoff/approach and cruise conditions are reviewed with emphasis on: (1) single and counterrotation model tests in the NASA Lewis 9 by 15 (low speed) and 8 by 6 (high speed) wind tunnels, and (2) full scale flight tests of a 9 ft (2.74 m) diameter single rotation wing mounted tractor and a 11.7 ft (3.57 m) diameter counterrotation aft mounted pusher propeller. Comparisons of model data projected to flight with full scale flight data show good agreement validating the scale model wind tunnel approach. Likewise, comparisons of measured and predicted noise level show excellent agreement for both single and counterrotation propellers. Progress in describing angle of attack and installation effects is also summarized. Finally, the aeroacoustic issues associated with ducted propellers (very high bypass fans) are discussed.

Electron transfer precedes ATP hydrolysis during nitrogenase catalysis

PubMed Central

Duval, Simon; Danyal, Karamatullah; Shaw, Sudipta; Lytle, Anna K.; Dean, Dennis R.; Hoffman, Brian M.; Antony, Edwin; Seefeldt, Lance C.

2013-01-01

The biological reduction of N2 to NH3 catalyzed by Mo-dependent nitrogenase requires at least eight rounds of a complex cycle of events associated with ATP-driven electron transfer (ET) from the Fe protein to the catalytic MoFe protein, with each ET coupled to the hydrolysis of two ATP molecules. Although steps within this cycle have been studied for decades, the nature of the coupling between ATP hydrolysis and ET, in particular the order of ET and ATP hydrolysis, has been elusive. Here, we have measured first-order rate constants for each key step in the reaction sequence, including direct measurement of the ATP hydrolysis rate constant: kATP = 70 s−1, 25 °C. Comparison of the rate constants establishes that the reaction sequence involves four sequential steps: (i) conformationally gated ET (kET = 140 s−1, 25 °C), (ii) ATP hydrolysis (kATP = 70 s−1, 25 °C), (iii) Phosphate release (kPi = 16 s−1, 25 °C), and (iv) Fe protein dissociation from the MoFe protein (kdiss = 6 s−1, 25 °C). These findings allow completion of the thermodynamic cycle undergone by the Fe protein, showing that the energy of ATP binding and protein–protein association drive ET, with subsequent ATP hydrolysis and Pi release causing dissociation of the complex between the Feox(ADP)2 protein and the reduced MoFe protein. PMID:24062462

ATP-sulfurylase, sulfur-compounds, and plant stress tolerance

PubMed Central

Anjum, Naser A.; Gill, Ritu; Kaushik, Manjeri; Hasanuzzaman, Mirza; Pereira, Eduarda; Ahmad, Iqbal; Tuteja, Narendra; Gill, Sarvajeet S.

2015-01-01

Sulfur (S) stands fourth in the list of major plant nutrients after N, P, and K. Sulfate (SO42-), a form of soil-S taken up by plant roots is metabolically inert. As the first committed step of S-assimilation, ATP-sulfurylase (ATP-S) catalyzes SO42--activation and yields activated high-energy compound adenosine-5′-phosphosulfate that is reduced to sulfide (S2-) and incorporated into cysteine (Cys). In turn, Cys acts as a precursor or donor of reduced S for a range of S-compounds such as methionine (Met), glutathione (GSH), homo-GSH (h-GSH), and phytochelatins (PCs). Among S-compounds, GSH, h-GSH, and PCs are known for their involvement in plant tolerance to varied abiotic stresses, Cys is a major component of GSH, h-GSH, and PCs; whereas, several key stress-metabolites such as ethylene, are controlled by Met through its first metabolite S-adenosylmethionine. With the major aim of briefly highlighting S-compound-mediated role of ATP-S in plant stress tolerance, this paper: (a) overviews ATP-S structure/chemistry and occurrence, (b) appraises recent literature available on ATP-S roles and regulations, and underlying mechanisms in plant abiotic and biotic stress tolerance, (c) summarizes ATP-S-intrinsic regulation by major S-compounds, and (d) highlights major open-questions in the present context. Future research in the current direction can be devised based on the discussion outcomes. PMID:25904923

Cerebrospinal fluid ATP metabolites in multiple sclerosis.

PubMed

Lazzarino, G; Amorini, A M; Eikelenboom, M J; Killestein, J; Belli, A; Di Pietro, V; Tavazzi, B; Barkhof, F; Polman, C H; Uitdehaag, B M J; Petzold, A

2010-05-01

Increased axonal energy demand and mitochondrial failure have been suggested as possible causes for axonal degeneration and disability in multiple sclerosis. Our objective was to test whether ATP depletion precedes clinical, imaging and biomarker evidence for axonal degeneration in multiple sclerosis. The method consisted of a longitudinal study which included 21 patients with multiple sclerosis. High performance liquid chromatography was used to quantify biomarkers of the ATP metabolism (oxypurines and purines) from the cerebrospinal fluid at baseline. The Expanded Disability Status Scale, MRI brain imaging measures for brain atrophy (ventricular and parenchymal fractions), and cerebrospinal fluid biomarkers for axonal damage (phosphorylated and hyperphosphorylated neurofilaments) were quantified at baseline and 3-year follow-up. Central ATP depletion (sum of ATP metabolites >19.7 micromol/litre) was followed by more severe progression of disability if compared to normal ATP metabolites (median 1.5 versus 0, p< 0.05). Baseline ATP metabolite levels correlated with change of Expanded Disability Status Scale in the pooled cohort (r= 0.66, p= 0.001) and subgroups (relapsing-remitting patients: r= 0.79, p< 0.05 and secondary progressive/primary progressive patients: r= 0.69, p< 0.01). There was no relationship between central ATP metabolites and either biomarker or MRI evidence for axonal degeneration. The data suggests that an increased energy demand in multiple sclerosis may cause a quantifiable degree of central ATP depletion. We speculate that the observed clinical disability may be related to depolarisation associated conduction block.

Responses of Rat P2X2 Receptors to Ultrashort Pulses of ATP Provide Insights into ATP Binding and Channel Gating

PubMed Central

Moffatt, Luciano; Hume, Richard I.

2007-01-01

To gain insight into the way that P2X2 receptors localized at synapses might function, we explored the properties of outside-out patches containing many of these channels as ATP was very rapidly applied and removed. Using a new method to calibrate the speed of exchange of solution over intact patches, we were able to reliably produce applications of ATP lasting <200 μs. For all concentrations of ATP, there was a delay of at least 80 μs between the time when ATP arrived at the receptor and the first detectable flow of inward current. In response to 200-μs pulses of ATP, the time constant of the rising phase of the current was ∼600 μs. Thus, most channel openings occurred when no free ATP was present. The current deactivated with a time constant of ∼60 ms. The amplitude of the peak response to a brief pulse of a saturating concentration of ATP was ∼70% of that obtained during a long application of the same concentration of ATP. Thus, ATP leaves fully liganded channels without producing an opening at least 30% of the time. Extensive kinetic modeling revealed three different schemes that fit the data well, a sequential model and two allosteric models. To account for the delay in opening at saturating ATP, it was necessary to incorporate an intermediate closed state into all three schemes. These kinetic properties indicate that responses to ATP at synapses that use homomeric P2X2 receptors would be expected to greatly outlast the duration of the synaptic ATP transient produced by a single presynaptic spike. Like NMDA receptors, P2X2 receptors provide the potential for complex patterns of synaptic integration over a time scale of hundreds of milliseconds. PMID:17664346

Study on Design of High Efficiency and Light Weight Composite Propeller Blade for a Regional Turboprop Aircraft

NASA Astrophysics Data System (ADS)

Kong, Changduk; Lee, Kyungsun

2013-03-01

In this study, aerodynamic and structural design of the composite propeller blade for a regional turboprop aircraft is performed. The thin and wide chord propeller blade of high speed turboprop aircraft should have proper strength and stiffness to carry various kinds of loads such as high aerodynamic bending and twisting moments and centrifugal forces. Therefore the skin-spar-foam sandwich structure using high strength and stiffness carbon/epoxy composite materials is used to improve the lightness. A specific design procedure is proposed in this work as follows; firstly the aerodynamic configuration design, which is acceptable for the design requirements, is carried out using the in-house code developed by authors, secondly the structure design loads are determined through the aerodynamic load case analysis, thirdly the spar flange and the skin are preliminarily sized by consideration of major bending moments and shear forces using both the netting rule and the rule of mixture, and finally, the stress analysis is performed to confirm the structural safety and stability using finite element analysis commercial code, MSC. NASTRAN/PATRAN. Furthermore the additional analysis is performed to confirm the structural safety due to bird strike impact on the blade during flight operation using a commercial code, ANSYS. To realize the proposed propeller design, the prototype blades are manufactured by the following procedure; the carbon/epoxy composite fabric prepregs are laid up for skin and spar on a mold using the hand lay-up method and consolidated with a proper temperature and vacuum in the oven. To finalize the structural design, the full-scale static structural test is performed under the simulated aerodynamic loads using 3 point loading method. From the experimental results, it is found that the designed blade has a good structural integrity, and the measured results agree well with the analytical results as well.

TCDD decreases ATP levels and increases reactive oxygen production through changes in mitochondrial F F{sub 1}-ATP synthase and ubiquinone

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shertzer, Howard G.; Genter, Mary Beth; Shen, Dongxiao

2006-12-15

Mitochondria generate ATP and participate in signal transduction and cellular pathology and/or cell death. TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) decreases hepatic ATP levels and generates mitochondrial oxidative DNA damage, which is exacerbated by increasing mitochondrial glutathione redox state and by inner membrane hyperpolarization. This study identifies mitochondrial targets of TCDD that initiate and sustain reactive oxygen production and decreased ATP levels. One week after treating mice with TCDD, liver ubiquinone (Q) levels were significantly decreased, while rates of succinoxidase and Q-cytochrome c oxidoreductase activities were increased. However, the expected increase in Q reduction state following TCDD treatment did not occur; instead, Q wasmore » more oxidized. These results could be explained by an ATP synthase defect, a premise supported by the unusual finding that TCDD lowers ATP/O ratios without concomitant changes in respiratory control ratios. Such results suggest either a futile cycle in ATP synthesis, or hydrolysis of newly synthesized ATP prior to release. The TCDD-mediated decrease in Q, concomitant with an increase in respiration, increases complex 3 redox cycling. This acts in concert with glutathione to increase membrane potential and reactive oxygen production. The proposed defect in ATP synthase explains both the greater respiratory rates and the lower tissue ATP levels.« less

Impact of age on exercise-induced ATP supply during supramaximal plantar flexion in humans

PubMed Central

Trinity, Joel D.; Hart, Corey R.; Kim, Seong-Eun; Groot, H. Jonathan; Fur, Yann Le; Sorensen, Jacob R.; Jeong, Eun-Kee; Richardson, Russell S.

2015-01-01

Currently, the physiological factors responsible for exercise intolerance and bioenergetic alterations with age are poorly understood due, at least in art, to the confounding effect of reduced physical activity in the elderly. Thus, in 40 healthy young (22 ± 2 yr) and old (74 ± 8 yr) activity-matched subjects, we assessed the impact of age on: 1) the relative contribution of the three major pathways of ATP synthesis (oxidative ATP synthesis, glycolysis, and the creatine kinase reaction) and 2) the ATP cost of contraction during high-intensity exercise. Specifically, during supramaximal plantar flexion (120% of maximal aerobic power), to stress the functional limits of the skeletal muscle energy systems, we used 31P-labeled magnetic resonance spectroscopy to assess metabolism. Although glycolytic activation was delayed in the old, ATP synthesis from the main energy pathways was not significantly different between groups. Similarly, the inferred peak rate of mitochondrial ATP synthesis was not significantly different between the young (25 ± 8 mM/min) and old (24 ± 6 mM/min). In contrast, the ATP cost of contraction was significantly elevated in the old compared with the young (5.1 ± 2.0 and 3.7 ± 1.7 mM·min−1·W−1, respectively; P < 0.05). Overall, these findings suggest that, when young and old subjects are activity matched, there is no evidence of age-related mitochondrial and glycolytic dysfunction. However, this study does confirm an abnormal elevation in exercise-induced skeletal muscle metabolic demand in the old that may contribute to the decline in exercise capacity with advancing age. PMID:26041112

ATP synthesis in Halobacterium saccharovorum: evidence that synthesis may be catalysed by an F0F1-ATP synthase

NASA Technical Reports Server (NTRS)

Hochstein, L. I.

1992-01-01

Halobacterium saccharovorum synthesized ATP in response to a pH shift from 8 to 6.2. Synthesis was inhibited by carbonyl cyanide m-chloro-phenylhydrazone, dicyclohexylcarbodiimide, and azide. Nitrate, an inhibitor of the membrane-bound ATPase previously isolated from this organism, did not inhibit ATP synthesis. N-Ethymaleimide, which also inhibited this ATPase, stimulated the production of ATP. These observations suggested that H. saccharovorum synthesized and hydrolysed ATP using different enzymes and that the vacuolar-like ATPase activity previously described in H. saccharovorum was an ATPase whose function is yet to be identified.

Bioanalytical Applications of Real-Time ATP Imaging Via Bioluminescence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gruenhagen, Jason Alan

The research discussed within involves the development of novel applications of real-time imaging of adenosine 5'-triphosphate (ATP). ATP was detected via bioluminescence and the firefly luciferase-catalyzed reaction of ATP and luciferin. The use of a microscope and an imaging detector allowed for spatially resolved quantitation of ATP release. Employing this method, applications in both biological and chemical systems were developed. First, the mechanism by which the compound 48/80 induces release of ATP from human umbilical vein endothelial cells (HUVECs) was investigated. Numerous enzyme activators and inhibitors were utilized to probe the second messenger systems involved in release. Compound 48/80 activatedmore » a G{sub q}-type protein to initiate ATP release from HUVECs. Ca 2+ imaging along with ATP imaging revealed that activation of phospholipase C and induction of intracellular Ca 2+ signaling were necessary for release of ATP. Furthermore, activation of protein kinase C inhibited the activity of phospholipase C and thus decreased the magnitude of ATP release. This novel release mechanism was compared to the existing theories of extracellular release of ATP. Bioluminescence imaging was also employed to examine the role of ATP in the field of neuroscience. The central nervous system (CNS) was dissected from the freshwater snail Lymnaea stagnalis. Electrophysiological experiments demonstrated that the neurons of the Lymnaea were not damaged by any of the components of the imaging solution. ATP was continuously released by the ganglia of the CNS for over eight hours and varied from ganglion to ganglion and within individual ganglia. Addition of the neurotransmitters K + and serotonin increased release of ATP in certain regions of the Lymnaea CNS. Finally, the ATP imaging technique was investigated for the study of drug release systems. MCM-41-type mesoporous nanospheres were loaded with ATP and end-capped with mercaptoethanol functionalized Cd

Inhibitors of the 5-lipoxygenase arachidonic acid pathway induce ATP release and ATP-dependent organic cation transport in macrophages.

PubMed

da Silva-Souza, Hercules Antônio; Lira, Maria Nathalia de; Costa-Junior, Helio Miranda; da Cruz, Cristiane Monteiro; Vasconcellos, Jorge Silvio Silva; Mendes, Anderson Nogueira; Pimenta-Reis, Gabriela; Alvarez, Cora Lilia; Faccioli, Lucia Helena; Serezani, Carlos Henrique; Schachter, Julieta; Persechini, Pedro Muanis

2014-07-01

We have previously described that arachidonic acid (AA)-5-lipoxygenase (5-LO) metabolism inhibitors such as NDGA and MK886, inhibit cell death by apoptosis, but not by necrosis, induced by extracellular ATP (ATPe) binding to P2X7 receptors in macrophages. ATPe binding to P2X7 also induces large cationic and anionic organic molecules uptake in these cells, a process that involves at least two distinct transport mechanisms: one for cations and another for anions. Here we show that inhibitors of the AA-5-LO pathway do not inhibit P2X7 receptors, as judged by the maintenance of the ATPe-induced uptake of fluorescent anionic dyes. In addition, we describe two new transport phenomena induced by these inhibitors in macrophages: a cation-selective uptake of fluorescent dyes and the release of ATP. The cation uptake requires secreted ATPe, but, differently from the P2X7/ATPe-induced phenomena, it is also present in macrophages derived from mice deficient in the P2X7 gene. Inhibitors of phospholipase A2 and of the AA-cyclooxygenase pathway did not induce the cation uptake. The uptake of non-organic cations was investigated by measuring the free intracellular Ca(2+) concentration ([Ca(2+)]i) by Fura-2 fluorescence. NDGA, but not MK886, induced an increase in [Ca(2+)]i. Chelating Ca(2+) ions in the extracellular medium suppressed the intracellular Ca(2+) signal without interfering in the uptake of cationic dyes. We conclude that inhibitors of the AA-5-LO pathway do not block P2X7 receptors, trigger the release of ATP, and induce an ATP-dependent uptake of organic cations by a Ca(2+)- and P2X7-independent transport mechanism in macrophages. Copyright © 2014 Elsevier B.V. All rights reserved.

Clusterin (Apolipoprotein J), a Molecular Chaperone That Facilitates Degradation of the Copper-ATPases ATP7A and ATP7B*

PubMed Central

Materia, Stephanie; Cater, Michael A.; Klomp, Leo W. J.; Mercer, Julian F. B.; La Fontaine, Sharon

2011-01-01

The copper-transporting P1B-type ATPases (Cu-ATPases) ATP7A and ATP7B are key regulators of physiological copper levels. They function to maintain intracellular copper homeostasis by delivering copper to secretory compartments and by trafficking toward the cell periphery to export excess copper. Mutations in the genes encoding ATP7A and ATP7B lead to copper deficiency and toxicity disorders, Menkes and Wilson diseases, respectively. This report describes the interaction between the Cu-ATPases and clusterin and demonstrates a chaperone-like role for clusterin in facilitating their degradation. Clusterin interacted with both ATP7A and ATP7B in mammalian cells. This interaction increased under conditions of oxidative stress and with mutations in ATP7B that led to its misfolding and mislocalization. A Wilson disease patient mutation (G85V) led to enhanced ATP7B turnover, which was further exacerbated when cells overexpressed clusterin. We demonstrated that clusterin-facilitated degradation of mutant ATP7B is likely to involve the lysosomal pathway. The knockdown and overexpression of clusterin increased and decreased, respectively, the Cu-ATPase-mediated copper export capacity of cells. These results highlight a new role for intracellular clusterin in mediating Cu-ATPase quality control and hence in the normal maintenance of copper homeostasis, and in promoting cell survival in the context of disease. Based on our findings, it is possible that variations in clusterin expression and function could contribute to the variable clinical expression of Menkes and Wilson diseases. PMID:21242307

Effect of Synthesized Propeller Vibration on Passenger Annoyance in a Turboprop Interior Noise Environment

NASA Technical Reports Server (NTRS)

Clevenson, S. A.

1982-01-01

The effect of synthesized propeller vibration on passenger annoyance to aircraft noise was investigated in passenger ride quality apparatus. Passenger reactions of annoyance to a wide range of potential turboprop interior noise environments were obtained under three simulated vibration conditions: no vibration, armrest vibration, and armrest plus cabin vibration. The noises, ranging from 71 to 95 dB(A) consisted of a turbulent boundary layer with a factorial combination of five blade passage frequencies (50 to 200 Hz), two harmonic roll offs, and three tone to noise ratios. Results indicate that passenger annoyance to noise in the presence of armrest vibration did not significantly change. However, those passengers exposed to cabin plus armrest vibration while being exposed to noise lower rating for the combined cabin vibration and noise environment compared with the rating for the noise along environment. This result is predicted by the ride quality model.

The AC-120: The advanced commercial transport

NASA Technical Reports Server (NTRS)

Duran, David; Griffin, Ernest; Mendoza, Saul; Nguyen, Son; Pickett, Tim; Noernberg, Clemm

1993-01-01

The main objective of this design was to fulfill a need for a new airplane to replace the aging 100 to 150 passenger, 1500 nautical mile range aircraft such as the Douglas DC9 and Boeing 737-100 airplanes. After researching the future aircraft market, conducting extensive trade studies, and analysis on different configurations, the AC-120 Advanced Commercial Transport final design was achieved. The AC-120's main design features include the incorporation of a three lifting surface configuration which is powered by two turboprop engines. The AC-120 is an economically sensitive aircraft which meets the new FM Stage Three noise requirements, and has lower NO(x) emissions than current turbofan powered airplanes. The AC-120 also improves on its contemporaries in passenger comfort, manufacturing, and operating cost.

Minoxidil opens mitochondrial K(ATP) channels and confers cardioprotection.

PubMed

Sato, Toshiaki; Li, Yulong; Saito, Tomoaki; Nakaya, Haruaki

2004-01-01

1. ATP-sensitive potassium channel in the mitochondrial inner membrane (mitoK(ATP) channel) rather than in the sarcolemma (sarcK(ATP) channel) appears to play an important role in cardioprotection. We examined the effect of minoxidil, a potent antihypertensive agent and hair growth stimulator, on sarcK(ATP) and mitoK(ATP) channels in guinea-pig ventricular myocytes. 2. Minoxidil activated a glybenclamide-sensitive sarcK(ATP) channel current in the whole-cell recording mode with an EC(50) of 182.6 microm. Minoxidil reversibly increased the flavoprotein oxidation, an index of mitoK(ATP) channel activity, in a concentration-dependent manner. The EC(50) for mitoK(ATP) channel activation was estimated to be 7.3 microm; this value was notably approximately 25-fold lower than that for sarcK(ATP) channel activation. 3. Minoxidil (10 microm) significantly attenuated the ouabain-induced increase of mitochondrial Ca(2+) concentration, which was measured by loading cells with rhod-2 fluorescence. Furthermore, pretreatment with minoxidil (10 microm) before 20-min no-flow ischaemia significantly improved the recovery of developed tension measured after 60 min of reperfusion in coronary perfused guinea-pig ventricular muscles. These cardioprotective effects of minoxidil were completely abolished by the mitoK(ATP) channel blocker 5-hydroxydecanoate (500 microm). 4. Our results indicate that minoxidil exerts a direct cardioprotective effect on heart muscle cells, an effect mediated by the selective activation of mitoK(ATP) channels.

Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus , a New Twist on ATP Formation

DOE PAGES

James, Kimberly L.; Ríos-Hernández, Luis A.; Wofford, Neil Q.; ...

2016-08-16

Syntrophus aciditrophicusis a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formate-consuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation byS. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome ofS. aciditrophicusleaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show thatS. aciditrophicususes AMP-forming, acetyl-CoA synthetase (Acs1)more » for ATP synthesis from acetyl-CoA.acs1mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, ofS. aciditrophicusgrown in pure culture and coculture. Cell extracts ofS. aciditrophicushad low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified fromS. aciditrophicusand recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) inS. aciditrophicuscells support the operation of Acs1 in the acetate-forming direction. Thus,S. aciditrophicushas a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase. We find bacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA.Syntrophus aciditrophicusapparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as

Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus , a New Twist on ATP Formation

DOE Office of Scientific and Technical Information (OSTI.GOV)

James, Kimberly L.; Ríos-Hernández, Luis A.; Wofford, Neil Q.

Syntrophus aciditrophicusis a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formate-consuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation byS. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome ofS. aciditrophicusleaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show thatS. aciditrophicususes AMP-forming, acetyl-CoA synthetase (Acs1)more » for ATP synthesis from acetyl-CoA.acs1mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, ofS. aciditrophicusgrown in pure culture and coculture. Cell extracts ofS. aciditrophicushad low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified fromS. aciditrophicusand recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) inS. aciditrophicuscells support the operation of Acs1 in the acetate-forming direction. Thus,S. aciditrophicushas a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase. We find bacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA.Syntrophus aciditrophicusapparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as

ATP depletion inhibits glucocorticoid-induced thymocyte apoptosis.

PubMed Central

Stefanelli, C; Bonavita, F; Stanic', I; Farruggia, G; Falcieri, E; Robuffo, I; Pignatti, C; Muscari, C; Rossoni, C; Guarnieri, C; Caldarera, C M

1997-01-01

In quiescent thymocytes, mitochondrial de-energization was not correlated to apoptotic death. In fact, thymocytes treated with oligomycin, a highly specific inhibitor of ATP synthase, alone or with atractyloside to block ATP translocation from the cytoplasm, were alive, even if their mitochondria were depolarized, as revealed by flow cytometry after Rhodamine 123 staining. Furthermore, oligomycin was a powerful inhibitor of apoptosis induced in rat thymocytes by dexamethasone and, to a lesser extent, by the calcium ionophore A23187 and etoposide, but was without effect when apoptosis was induced by staurosporine, and increased cell death in mitogen-treated thymocytes. The inhibition of apoptosis was confirmed by morphological criteria, inhibition of inter-nucleosomal DNA fragmentation and inhibition of the loss of membrane integrity. The anti-apoptotic effect of oligomycin in cells treated with A23187 or etoposide was correlated to the inhibition of protein synthesis, while inhibition of apoptosis induced by dexamethasone, already evident at an oligomycin concentration of 10 ng/ml, was instead strictly correlated to the effect exerted on the cellular ATP level. Thymocyte apoptosis triggered by dexamethasone was blocked or delayed by inhibitors of respiratory-chain uncouplers, inhibitors of ATP synthase and antioxidants: a lasting protection from dexamethasone-induced apoptosis was always correlated to a drastic and rapid reduction in ATP level (31-35% of control), while a delay in the death process was characterized by a moderate decrease in ATP (73-82% of control). Oligomycin inhibited the specific binding of radioactive corticosteroid to thymocyte nuclei, confirming the inhibitory effect of ATP depletion on glucocorticoid binding and suggesting that ATP depletion is a common mediator of the anti-apoptotic action of different effectors in glucocorticoid-induced apoptosis. In conclusion, the reported data indicate that ATP may act as a cellular modulator of some

Stretch-induced Ca2+ independent ATP release in hippocampal astrocytes.

PubMed

Xiong, Yingfei; Teng, Sasa; Zheng, Lianghong; Sun, Suhua; Li, Jie; Guo, Ning; Li, Mingli; Wang, Li; Zhu, Feipeng; Wang, Changhe; Rao, Zhiren; Zhou, Zhuan

2018-02-28

Similar to neurons, astrocytes actively participate in synaptic transmission via releasing gliotransmitters. The Ca 2+ -dependent release of gliotransmitters includes glutamate and ATP. Following an 'on-cell-like' mechanical stimulus to a single astrocyte, Ca 2+ independent single, large, non-quantal, ATP release occurs. Astrocytic ATP release is inhibited by either selective antagonist treatment or genetic knockdown of P2X7 receptor channels. Our work suggests that ATP can be released from astrocytes via two independent pathways in hippocampal astrocytes; in addition to the known Ca 2+ -dependent vesicular release, larger non-quantal ATP release depends on P2X7 channels following mechanical stretch. Astrocytic ATP release is essential for brain functions such as synaptic long-term potentiation for learning and memory. However, whether and how ATP is released via exocytosis remains hotly debated. All previous studies of non-vesicular ATP release have used indirect assays. By contrast, two recent studies report vesicular ATP release using more direct assays. In the present study, using patch clamped 'ATP-sniffer cells', we re-investigated astrocytic ATP release at single-vesicle resolution in hippocampal astrocytes. Following an 'on-cell-like' mechanical stimulus of a single astrocyte, a Ca 2+ independent single large non-quantal ATP release occurred, in contrast to the Ca 2+ -dependent multiple small quantal ATP release in a chromaffin cell. The mechanical stimulation-induced ATP release from an astrocyte was inhibited by either exposure to a selective antagonist or genetic knockdown of P2X7 receptor channels. Functional P2X7 channels were expressed in astrocytes in hippocampal brain slices. Thus, in addition to small quantal ATP release, larger non-quantal ATP release depends on P2X7 channels in astrocytes. © 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

ATP7A is a novel target of retinoic acid receptor β2 in neuroblastoma cells

PubMed Central

Bohlken, A; Cheung, B B; Bell, J L; Koach, J; Smith, S; Sekyere, E; Thomas, W; Norris, M; Haber, M; Lovejoy, D B; Richardson, D R; Marshall, G M

2009-01-01

Increased retinoic acid receptor β (RARβ2) gene expression is a hallmark of cancer cell responsiveness to retinoid anticancer effects. Moreover, low basal or induced RARβ2 expression is a common feature of many human cancers, suggesting that RARβ2 may act as a tumour suppressor gene in the absence of supplemented retinoid. We have previously shown that low RARβ2 expression is a feature of advanced neuroblastoma. Here, we demonstrate that the ABC domain of the RARβ2 protein alone was sufficient for the growth inhibitory effects of RARβ2 on neuroblastoma cells. ATP7A, the copper efflux pump, is a retinoid-responsive gene, was upregulated by ectopic overexpression of RARβ2. The ectopic overexpression of the RARβ2 ABC domain was sufficient to induce ATP7A expression, whereas, RARβ2 siRNA blocked the induction of ATP7A expression in retinoid-treated neuroblastoma cells. Forced downregulation of ATP7A reduced copper efflux and increased viability of retinoid-treated neuroblastoma cells. Copper supplementation enhanced cell growth and reduced retinoid-responsiveness, whereas copper chelation reduced the viability and proliferative capacity. Taken together, our data demonstrates ATP7A expression is regulated by retinoic acid receptor β and it has effects on intracellular copper levels, revealing a link between the anticancer action of retinoids and copper metabolism. PMID:19127267

Kinetic mechanism of the dimeric ATP sulfurylase from plants

PubMed Central

Ravilious, Geoffrey E.; Herrmann, Jonathan; Goo Lee, Soon; Westfall, Corey S.; Jez, Joseph M.

2013-01-01

In plants, sulfur must be obtained from the environment and assimilated into usable forms for metabolism. ATP sulfurylase catalyses the thermodynamically unfavourable formation of a mixed phosphosulfate anhydride in APS (adenosine 5′-phosphosulfate) from ATP and sulfate as the first committed step of sulfur assimilation in plants. In contrast to the multi-functional, allosterically regulated ATP sulfurylases from bacteria, fungi and mammals, the plant enzyme functions as a mono-functional, non-allosteric homodimer. Owing to these differences, here we examine the kinetic mechanism of soybean ATP sulfurylase [GmATPS1 (Glycine max (soybean) ATP sulfurylase isoform 1)]. For the forward reaction (APS synthesis), initial velocity methods indicate a single-displacement mechanism. Dead-end inhibition studies with chlorate showed competitive inhibition versus sulfate and non-competitive inhibition versus APS. Initial velocity studies of the reverse reaction (ATP synthesis) demonstrate a sequential mechanism with global fitting analysis suggesting an ordered binding of substrates. ITC (isothermal titration calorimetry) showed tight binding of APS to GmATPS1. In contrast, binding of PPi (pyrophosphate) to GmATPS1 was not detected, although titration of the E•APS complex with PPi in the absence of magnesium displayed ternary complex formation. These results suggest a kinetic mechanism in which ATP and APS are the first substrates bound in the forward and reverse reactions, respectively. PMID:23789618

ATP13A2 variability in Parkinson disease

PubMed Central

Vilariño-Güell, Carles; Soto, Alexandra I.; Lincoln, Sarah J.; Yahmed, Samia Ben; Kefi, Mounir; Heckman, Michael G.; Hulihan, Mary M.; Chai, Hua; Diehl, Nancy N.; Amouri, Rim; Rajput, Alex; Mash, Deborah C.; Dickson, Dennis W.; Middleton, Lefkos T.; Gibson, Rachel A.; Hentati, Faycal; Farrer, Matthew J.

2008-01-01

Recessively inherited mutations in ATP13A2 result in Kufor-Rakeb syndrome, whereas genetic variability and elevated ATP13A2 expression have been implicated in Parkinson disease (PD). Given this background, ATP13A2 was comprehensively assessed to support or refute its contribution to PD. Sequencing of ATP13A2 exons and intron-exon boundaries was performed in 89 probands with familial parkinsonism from Tunisia. The segregation of mutations with parkinsonism was subsequently assessed within pedigrees. The frequency of genetic variants and evidence for association was also examined in 240 patients with non-familial PD and 372 healthy controls. ATP13A2 mRNA expression was also quantified in brain tissues from 38 patients with non-familial PD and 38 healthy subjects from the US. Sequencing analysis revealed 37 new variants; seven missense, six silent and 24 that were noncoding. However, no single ATP13A2 mutation segregated with familial parkinsonism in either a dominant or recessive manner. Four markers showed marginal association with non-familial PD, prior to correction for multiple testing. ATP13A2 mRNA expression was marginally decreased in PD brains compared with tissue from control subjects. In conclusion, neither ATP13A2 genetic variability nor quantitative gene expression in brain appears to contribute to familial parkinsonism or non-familial PD. PMID:19085912

An RNA motif that binds ATP

NASA Technical Reports Server (NTRS)

Sassanfar, M.; Szostak, J. W.

1993-01-01

RNAs that contain specific high-affinity binding sites for small molecule ligands immobilized on a solid support are present at a frequency of roughly one in 10(10)-10(11) in pools of random sequence RNA molecules. Here we describe a new in vitro selection procedure designed to ensure the isolation of RNAs that bind the ligand of interest in solution as well as on a solid support. We have used this method to isolate a remarkably small RNA motif that binds ATP, a substrate in numerous biological reactions and the universal biological high-energy intermediate. The selected ATP-binding RNAs contain a consensus sequence, embedded in a common secondary structure. The binding properties of ATP analogues and modified RNAs show that the binding interaction is characterized by a large number of close contacts between the ATP and RNA, and by a change in the conformation of the RNA.

A High-Throughput TNP-ATP Displacement Assay for Screening Inhibitors of ATP-Binding in Bacterial Histidine Kinases

PubMed Central

Guarnieri, Michael T.; Blagg, Brian S. J.

2011-01-01

Abstract Bacterial histidine kinases (HK) are members of the GHKL superfamily, which share a unique adenosine triphosphate (ATP)-binding Bergerat fold. Our previous studies have shown that Gyrase, Hsp90, MutL (GHL) inhibitors bind to the ATP-binding pocket of HK and may provide lead compounds for the design of novel antibiotics targeting these kinases. In this article, we developed a competition assay using the fluorescent ATP analog, 2′,3′-O-(2,4,6-trinitrophenyl) adenosine 5′-triphosphate. The method can be used for high-throughput screening of compound libraries targeting HKs or other ATP-binding proteins. We utilized the assay to screen a library of GHL inhibitors targeting the bacterial HK PhoQ, and discuss the applications of the 2′,3′-O-(2,4,6-trinitrophenyl) adenosine 5′-triphosphate competition assay beyond GHKL inhibitor screening. PMID:21050069

Theoretical studies of the ATP hydrolysis mechanism of myosin.

PubMed

Okimoto, N; Yamanaka, K; Ueno, J; Hata, M; Hoshino, T; Tsuda, M

2001-11-01

The ATP hydrolysis mechanism of myosin was studied using quantum chemical (QM) and molecular dynamics calculations. The initial model compound for QM calculations was constructed on the basis of the energy-minimized structure of the myosin(S1dc)-ATP complex, which was determined by molecular mechanics calculations. The result of QM calculations suggested that the ATP hydrolysis mechanism of myosin consists of a single elementary reaction in which a water molecule nucleophilically attacked gamma-phosphorus of ATP. In addition, we performed molecular dynamics simulations of the initial and final states of the ATP hydrolysis reaction, that is, the myosin-ATP and myosin-ADP.Pi complexes. These calculations revealed roles of several amino acid residues (Lys185, Thr186, Ser237, Arg238, and Glu459) in the ATPase pocket. Lys185 maintains the conformation of beta- and gamma-phosphate groups of ATP by forming the hydrogen bonds. Thr186 and Ser237 are coordinated to a Mg(2+) ion, which interacts with the phosphates of ATP and therefore contributes to the stabilization of the ATP structure. Arg238 and Glu459, which consisted of the gate of the ATPase pocket, retain the water molecule acting on the hydrolysis at the appropriate position for initiating the hydrolysis.

ATP and AMP Mutually Influence Their Interaction with the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) at Separate Binding Sites*

PubMed Central

Randak, Christoph O.; Dong, Qian; Ver Heul, Amanda R.; Elcock, Adrian H.; Welsh, Michael J.

2013-01-01

Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel in the ATP-binding cassette (ABC) transporter protein family. In the presence of ATP and physiologically relevant concentrations of AMP, CFTR exhibits adenylate kinase activity (ATP + AMP ⇆ 2 ADP). Previous studies suggested that the interaction of nucleotide triphosphate with CFTR at ATP-binding site 2 is required for this activity. Two other ABC proteins, Rad50 and a structural maintenance of chromosome protein, also have adenylate kinase activity. All three ABC adenylate kinases bind and hydrolyze ATP in the absence of other nucleotides. However, little is known about how an ABC adenylate kinase interacts with ATP and AMP when both are present. Based on data from non-ABC adenylate kinases, we hypothesized that ATP and AMP mutually influence their interaction with CFTR at separate binding sites. We further hypothesized that only one of the two CFTR ATP-binding sites is involved in the adenylate kinase reaction. We found that 8-azidoadenosine 5′-triphosphate (8-N3-ATP) and 8-azidoadenosine 5′-monophosphate (8-N3-AMP) photolabeled separate sites in CFTR. Labeling of the AMP-binding site with 8-N3-AMP required the presence of ATP. Conversely, AMP enhanced photolabeling with 8-N3-ATP at ATP-binding site 2. The adenylate kinase active center probe P1,P5-di(adenosine-5′) pentaphosphate interacted simultaneously with an AMP-binding site and ATP-binding site 2. These results show that ATP and AMP interact with separate binding sites but mutually influence their interaction with the ABC adenylate kinase CFTR. They further indicate that the active center of the adenylate kinase comprises ATP-binding site 2. PMID:23921386

Torque generation mechanism of ATP synthase

NASA Astrophysics Data System (ADS)

Miller, John; Maric, Sladjana; Scoppa, M.; Cheung, M.

2010-03-01

ATP synthase is a rotary motor that produces adenosine triphosphate (ATP), the chemical currency of life. Our proposed electric field driven torque (EFT) model of FoF1-ATP synthase describes how torque, which scales with the number of c-ring proton binding sites, is generated by the proton motive force (pmf) across the mitochondrial inner membrane. When Fo is coupled to F1, the model predicts a critical pmf to drive ATP production. In order to fully understand how the electric field resulting from the pmf drives the c-ring to rotate, it is important to examine the charge distributions in the protonated c-ring and a-subunit containing the proton channels. Our calculations use a self-consistent field approach based on a refinement of reported structural data. The results reveal changes in pKa for key residues on the a-subunit and c-ring, as well as titration curves and protonation state energy diagrams. Health implications will be briefly discussed.

ATP-dependent chromatin remodeling in T cells

PubMed Central

Wurster, Andrea L.; Pazin, Michael J.

2012-01-01

One of the best studied systems for mammalian chromatin remodeling is transcriptional regulation during T cell development. The variety of these studies have led to important findings in T cell gene regulation and cell fate determination. Importantly, these findings have also advanced our knowledge of the function of remodeling enzymes in mammalian gene regulation. In this review, first we briefly present biochemical/cell-free analysis of 3 types of ATP dependent remodeling enzymes (SWI/SNF, Mi2, and ISWI), to construct an intellectual framework to understand how these enzymes might be working. Second, we compare and contrast the function of these enzymes, during early (thymic) and late (peripheral) T cell development. Finally, we examine some of the gaps in our present understanding. PMID:21999456

ATP-dependent chromatin remodeling in T cells.

PubMed

Wurster, Andrea L; Pazin, Michael J

2012-02-01

One of the best studied systems for mammalian chromatin remodeling is transcriptional regulation during T cell development. The variety of these studies have led to important findings in T cell gene regulation and cell fate determination. Importantly, these findings have also advanced our knowledge of the function of remodeling enzymes in mammalian gene regulation. First we briefly present biochemical and cell-free analysis of 3 types of ATP dependent remodeling enzymes (SWI/SNF, Mi2, and ISWI) to construct an intellectual framework to understand how these enzymes might be working. Second, we compare and contrast the function of these enzymes during early (thymic) and late (peripheral) T cell development. Finally, we examine some of the gaps in our present understanding.

Dynamic Regulation of Cell Volume and Extracellular ATP of Human Erythrocytes

PubMed Central

Leal Denis, M. Florencia; Alvarez, H. Ariel; Lauri, Natalia; Alvarez, Cora L.; Chara, Osvaldo; Schwarzbaum, Pablo J.

2016-01-01

Introduction The peptide mastoparan 7 (MST7) triggered in human erythrocytes (rbcs) the release of ATP and swelling. Since swelling is a well-known inducer of ATP release, and extracellular (ATPe), interacting with P (purinergic) receptors, can affect cell volume (Vr), we explored the dynamic regulation between Vr and ATPe. Methods and Treatments We made a quantitative assessment of MST7-dependent kinetics of Vr and of [ATPe], both in the absence and presence of blockers of ATP efflux, swelling and P receptors. Results In rbcs 10 μM MST7 promoted acute, strongly correlated changes in [ATPe] and Vr. Whereas MST7 induced increases of 10% in Vr and 190 nM in [ATPe], blocking swelling in a hyperosmotic medium + MST7 reduced [ATPe] by 40%. Pre-incubation of rbcs with 10 μM of either carbenoxolone or probenecid, two inhibitors of the ATP conduit pannexin 1, reduced [ATPe] by 40–50% and swelling by 40–60%, while in the presence of 80 U/mL apyrase, an ATPe scavenger, cell swelling was prevented. While exposure to 10 μM NF110, a blocker of ATP-P2X receptors mediating sodium influx, reduced [ATPe] by 48%, and swelling by 80%, incubation of cells in sodium free medium reduced swelling by 92%. Analysis and Discussion Results were analyzed by means of a mathematical model where ATPe kinetics and Vr kinetics were mutually regulated. Model dependent fit to experimental data showed that, upon MST7 exposure, ATP efflux required a fast 1960-fold increase of ATP permeability, mediated by two kinetically different conduits, both of which were activated by swelling and inactivated by time. Both experimental and theoretical results suggest that, following MST7 exposure, ATP is released via two conduits, one of which is mediated by pannexin 1. The accumulated ATPe activates P2X receptors, followed by sodium influx, resulting in cell swelling, which in turn further activates ATP release. Thus swelling and P2X receptors constitute essential components of a positive feedback loop

ATP Synthase Diseases of Mitochondrial Genetic Origin

PubMed Central

Dautant, Alain; Meier, Thomas; Hahn, Alexander; Tribouillard-Tanvier, Déborah; di Rago, Jean-Paul; Kucharczyk, Roza

2018-01-01

Devastating human neuromuscular disorders have been associated to defects in the ATP synthase. This enzyme is found in the inner mitochondrial membrane and catalyzes the last step in oxidative phosphorylation, which provides aerobic eukaryotes with ATP. With the advent of structures of complete ATP synthases, and the availability of genetically approachable systems such as the yeast Saccharomyces cerevisiae, we can begin to understand these molecular machines and their associated defects at the molecular level. In this review, we describe what is known about the clinical syndromes induced by 58 different mutations found in the mitochondrial genes encoding membrane subunits 8 and a of ATP synthase, and evaluate their functional consequences with respect to recently described cryo-EM structures. PMID:29670542

Power Stroke Angular Velocity Profiles of Archaeal A-ATP Synthase Versus Thermophilic and Mesophilic F-ATP Synthase Molecular Motors*

PubMed Central

Sielaff, Hendrik; Martin, James; Singh, Dhirendra; Biuković, Goran; Grüber, Gerhard; Frasch, Wayne D.

2016-01-01

The angular velocities of ATPase-dependent power strokes as a function of the rotational position for the A-type molecular motor A3B3DF, from the Methanosarcina mazei Gö1 A-ATP synthase, and the thermophilic motor α3β3γ, from Geobacillus stearothermophilus (formerly known as Bacillus PS3) F-ATP synthase, are resolved at 5 μs resolution for the first time. Unexpectedly, the angular velocity profile of the A-type was closely similar in the angular positions of accelerations and decelerations to the profiles of the evolutionarily distant F-type motors of thermophilic and mesophilic origins, and they differ only in the magnitude of their velocities. M. mazei A3B3DF power strokes occurred in 120° steps at saturating ATP concentrations like the F-type motors. However, because ATP-binding dwells did not interrupt the 120° steps at limiting ATP, ATP binding to A3B3DF must occur during the catalytic dwell. Elevated concentrations of ADP did not increase dwells occurring 40° after the catalytic dwell. In F-type motors, elevated ADP induces dwells 40° after the catalytic dwell and slows the overall velocity. The similarities in these power stroke profiles are consistent with a common rotational mechanism for A-type and F-type rotary motors, in which the angular velocity is limited by the rotary position at which ATP binding occurs and by the drag imposed on the axle as it rotates within the ring of stator subunits. PMID:27729450

Hypophosphatemia promotes lower rates of muscle ATP synthesis.

PubMed

Pesta, Dominik H; Tsirigotis, Dimitrios N; Befroy, Douglas E; Caballero, Daniel; Jurczak, Michael J; Rahimi, Yasmeen; Cline, Gary W; Dufour, Sylvie; Birkenfeld, Andreas L; Rothman, Douglas L; Carpenter, Thomas O; Insogna, Karl; Petersen, Kitt Falk; Bergwitz, Clemens; Shulman, Gerald I

2016-10-01

Hypophosphatemia can lead to muscle weakness and respiratory and heart failure, but the mechanism is unknown. To address this question, we noninvasively assessed rates of muscle ATP synthesis in hypophosphatemic mice by using in vivo saturation transfer [ 31 P]-magnetic resonance spectroscopy. By using this approach, we found that basal and insulin-stimulated rates of muscle ATP synthetic flux (V ATP ) and plasma inorganic phosphate (P i ) were reduced by 50% in mice with diet-induced hypophosphatemia as well as in sodium-dependent P i transporter solute carrier family 34, member 1 (NaPi2a)-knockout (NaPi2a -/- ) mice compared with their wild-type littermate controls. Rates of V ATP normalized in both hypophosphatemic groups after restoring plasma P i concentrations. Furthermore, V ATP was directly related to cellular and mitochondrial P i uptake in L6 and RC13 rodent myocytes and isolated muscle mitochondria. Similar findings were observed in a patient with chronic hypophosphatemia as a result of a mutation in SLC34A3 who had a 50% reduction in both serum P i content and muscle V ATP After oral P i repletion and normalization of serum P i levels, muscle V ATP completely normalized in the patient. Taken together, these data support the hypothesis that decreased muscle ATP synthesis, in part, may be caused by low blood P i concentrations, which may explain some aspects of muscle weakness observed in patients with hypophosphatemia.-Pesta, D. H., Tsirigotis, D. N., Befroy, D. E., Caballero, D., Jurczak, M. J., Rahimi, Y., Cline, G. W., Dufour, S., Birkenfeld, A. L., Rothman, D. L., Carpenter, T. O., Insogna, K., Petersen, K. F., Bergwitz, C., Shulman, G. I. Hypophosphatemia promotes lower rates of muscle ATP synthesis. © The Author(s).

Lysosomal Storage of Subunit c of Mitochondrial ATP Synthase in Brain-Specific Atp13a2-Deficient Mice.

PubMed

Sato, Shigeto; Koike, Masato; Funayama, Manabu; Ezaki, Junji; Fukuda, Takahiro; Ueno, Takashi; Uchiyama, Yasuo; Hattori, Nobutaka

2016-12-01

Kufor-Rakeb syndrome (KRS) is an autosomal recessive form of early-onset parkinsonism linked to the PARK9 locus. The causative gene for KRS is Atp13a2, which encodes a lysosomal type 5 P-type ATPase. We recently showed that KRS/PARK9-linked mutations lead to several lysosomal alterations, including reduced proteolytic processing of cathepsin D in vitro. However, it remains unknown how deficiency of Atp13a2 is connected to lysosomal impairments. To address this issue, we analyzed brain tissues of Atp13a2 conditional-knockout mice, which exhibited characteristic features of neuronal ceroid lipofuscinosis, including accumulation of lipofuscin positive for subunit c of mitochondrial ATP synthase, suggesting that a common pathogenic mechanism underlies both neuronal ceroid lipofuscinosis and Parkinson disease. Copyright © 2016 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

The importance of ATP-related compounds for the freshness and flavor of post-mortem fish and shellfish muscle: A review.

PubMed

Hong, Hui; Regenstein, Joe M; Luo, Yongkang

2017-06-13

ATP degradation is one of the most important biochemical changes in the post-mortem muscle of fish and shellfish. This process has long been recognized as an accurate way to evaluate freshness of fish and shellfish product. This review updates and condenses the overall history and recent advances in understanding the role of ATP-related compounds in post-mortem fish and shellfish muscle including a discussion of key analytical methods, their use as a freshness indicator, their roles in flavor enhancement, the factors affecting their transitions, and the possible mechanisms responsible for their impact on flavor and freshness. Moreover, some challenges and future directions for research regarding ATP-related compounds in fish and shellfish flavor and freshness are presented. With increasing consumer demands for fresh products with extended shelf life, understanding the relationships between ATP-related compounds and their involvement in the freshness and umami taste is a prerequisite for assuring the high quality of fish and shellfish.

Binding of ATP by pertussis toxin and isolated toxin subunits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hausman, S.Z.; Manclark, C.R.; Burns, D.L.

1990-07-03

The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of ({sup 3}H)ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of ({sup 3}H)ATP to pertussis toxin in a competitive manner;more » however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of ({sup 3}H)ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site.« less

Identification of a new Mpl-interacting protein, Atp5d.

PubMed

Liu, Hongyan; Zhao, Zhenhu; Zhong, Yuxu; Shan, Yajun; Sun, Xiaohong; Mao, Bingzhi; Cong, Yuwen

2014-06-01

Thrombopoietin (TPO) can regulate hematopoiesis and megakaryopoiesis via activation of its receptor, c-Mpl, and multiple downstream signal transduction pathways. Using the cytoplasmic domain of Mpl as bait, we performed yeast two-hybrid screening, and found that the protein Atp5d might associate with Mpl. Atp5d is known as the δ subunit of mitochondrial ATP synthase, but little is known about the function of dissociative Atp5d. The interaction between Mpl and Atp5d was confirmed by the yeast two-hybrid system, mammalian two-hybrid assay, pull-down experiment, and co-immunoprecipitation study in vivo and in vitro. An additional immunofluorescence assay showed that the two proteins can colocalize along the plasma membrane in the cytoplasm. Using the yeast two-hybrid system, we tested a series of cytoplasmic truncated mutations for their ability to bind Atp5d and found an association between Atp5d and the Aa98-113 domain of Mpl. The dissociation of Atp5d from Mpl after TPO stimulation suggests that Atp5d may be a new component of TPO signaling.

Evidence that Na+/H+ exchanger 1 is an ATP-binding protein.

PubMed

Shimada-Shimizu, Naoko; Hisamitsu, Takashi; Nakamura, Tomoe Y; Wakabayashi, Shigeo

2013-03-01

Na(+)/H(+) exchanger (NHE) 1 is a member of the solute carrier superfamily, which regulates intracellular ionic homeostasis. NHE1 is known to require cellular ATP for its activity, despite there being no requirement for energy input from ATP hydrolysis. In this study, we investigated whether NHE1 is an ATP-binding protein. We designed a baculovirus vector carrying both epitope-tagged NHE1 and its cytosolic subunit CHP1, and expressed the functional NHE1-CHP1 complex on the surface of Sf9 insect cells. Using the purified complex protein consisting of NHE1 and CHP1 from Sf9 cells, we examined a photoaffinity labeling reaction with 8-azido-ATP-biotin. UV irradiation promoted the incorporation of 8-azido-ATP into NHE1, but not into CHP1, with an apparent Kd of 29.1 µM in the presence of Mg(2+). The nonlabeled nucleotides ATP, GTP, TTP and CTP all inhibited this crosslinking. However, ATP had the strongest inhibitory effect, with an apparent inhibition constant (IC50) for ATP of 2.2 mM, close to the ATP concentration giving the half-maximal activation of NHE1 activity. Importantly, crosslinking was more strongly inhibited by ATP than by ADP, suggesting that ATP is dissociated from NHE1 upon ATP hydrolysis. Limited proteolysis with thrombin and deletion mutant analysis revealed that the 8-azido-ATP-binding site is within the C-terminal cytoplasmic domain of NHE1. Equilibrium dialysis with NHE1-derived peptides provided evidence that ATP directly binds to the proximal cytoplasmic region (Gly542-Pro598), which is critical for ATP-dependent regulation of NHE1. These findings suggest that NHE1 is an ATP-binding transporter. Thus, ATP may serve as a direct activator of NHE1. © 2013 The Authors Journal compilation © 2013 FEBS.

An advanced pitch change mechanism incorporating a hybrid traction drive

NASA Technical Reports Server (NTRS)

Steinetz, B. M.; Sargisson, D. F.; White, G.; Loewenthal, S. H.

1984-01-01

A design of a propeller pitch control mechanism is described that meets the demanding requirements of a high-power, advanced turboprop. In this application, blade twisting moment torque can be comparable to that of the main reduction gearbox output: precise pitch control, reliability and compactness are all at a premium. A key element in the design is a compact, high-ratio hybrid traction drive which offers low torque ripple and high torsional stiffness. The traction drive couples a high speed electric motor/alternator unit to a ball screw that actuates the blade control links. The technical merits of this arrangement and the performance characteristics of the traction drive are discussed. Comparisons are made to the more conventional pitch control mechanisms.

In-flight source noise of an advanced full-scale single-rotation propeller

NASA Technical Reports Server (NTRS)

Woodward, Richard P.; Loffler, Irvin J.

1991-01-01

Flight tests to define the far-field tone source at cruise conditions have been completed on the full-scale SR-7L advanced turboprop, which was installed on the left wing of a Gulfstream II aircraft. These measurements defined source levels for input into long-distance propagation models to predict en route noise. Infight data were taken for seven test cases. The sideline directivities measured showed expected maximum levels near 105 deg from the propeller upstream axis. However, azimuthal directivities based on the maximum observed sideline tone levels showed highest levels below the aircraft. The tone level reduction associated with reductions in propeller tip speed is shown to be more significant in the horizontal plane than below the aircraft.

Power Stroke Angular Velocity Profiles of Archaeal A-ATP Synthase Versus Thermophilic and Mesophilic F-ATP Synthase Molecular Motors.

PubMed

Sielaff, Hendrik; Martin, James; Singh, Dhirendra; Biuković, Goran; Grüber, Gerhard; Frasch, Wayne D

2016-12-02

The angular velocities of ATPase-dependent power strokes as a function of the rotational position for the A-type molecular motor A 3 B 3 DF, from the Methanosarcina mazei Gö1 A-ATP synthase, and the thermophilic motor α 3 β 3 γ, from Geobacillus stearothermophilus (formerly known as Bacillus PS3) F-ATP synthase, are resolved at 5 μs resolution for the first time. Unexpectedly, the angular velocity profile of the A-type was closely similar in the angular positions of accelerations and decelerations to the profiles of the evolutionarily distant F-type motors of thermophilic and mesophilic origins, and they differ only in the magnitude of their velocities. M. mazei A 3 B 3 DF power strokes occurred in 120° steps at saturating ATP concentrations like the F-type motors. However, because ATP-binding dwells did not interrupt the 120° steps at limiting ATP, ATP binding to A 3 B 3 DF must occur during the catalytic dwell. Elevated concentrations of ADP did not increase dwells occurring 40° after the catalytic dwell. In F-type motors, elevated ADP induces dwells 40° after the catalytic dwell and slows the overall velocity. The similarities in these power stroke profiles are consistent with a common rotational mechanism for A-type and F-type rotary motors, in which the angular velocity is limited by the rotary position at which ATP binding occurs and by the drag imposed on the axle as it rotates within the ring of stator subunits. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Critical role of ATP-induced ATP release for Ca2+ signaling in nonsensory cell networks of the developing cochlea

PubMed Central

Ceriani, Federico; Pozzan, Tullio; Mammano, Fabio

2016-01-01

Spatially and temporally coordinated variations of the cytosolic free calcium concentration ([Ca2+]c) play a crucial role in a variety of tissues. In the developing sensory epithelium of the mammalian cochlea, elevation of extracellular adenosine trisphosphate concentration ([ATP]e) triggers [Ca2+]c oscillations and propagation of intercellular inositol 1,4,5-trisphosphate (IP3)-dependent Ca2+ waves. What remains uncertain is the relative contribution of gap junction channels and connexin hemichannels to these fundamental mechanisms, defects in which impair hearing acquisition. Another related open question is whether [Ca2+]c oscillations require oscillations of the cytosolic IP3 concentration ([IP3]c) in this system. To address these issues, we performed Ca2+ imaging experiments in the lesser epithelial ridge of the mouse cochlea around postnatal day 5 and constructed a computational model in quantitative adherence to experimental data. Our results indicate that [Ca2+]c oscillations are governed by Hopf-type bifurcations within the experimental range of [ATP]e and do not require [IP3]c oscillations. The model replicates accurately the spatial extent and propagation speed of intercellular Ca2+ waves and predicts that ATP-induced ATP release is the primary mechanism underlying intercellular propagation of Ca2+ signals. The model also uncovers a discontinuous transition from propagating regimes (intercellular Ca2+ wave speed > 11 μm⋅s−1) to propagation failure (speed = 0), which occurs upon lowering the maximal ATP release rate below a minimal threshold value. The approach presented here overcomes major limitations due to lack of specific connexin channel inhibitors and can be extended to other coupled cellular systems. PMID:27807138

Numerical modelling of flow and heat transfer in the rotating disc cavities of a turboprop engine.

PubMed

Faragher, J; Ooi, A

2001-05-01

A numerical analysis of the flow and heat transfer in the cavity between two co-rotating discs with axial inlet and radial outflow of fluid, a configuration common in gas turbine engines, is described. The results are compared with the experimental data of Northrop and Owen. The effectiveness of the k-epsilon turbulence model with the two-layer zonal model for near-wall treatment of Chen and Patel is tested for this type of flow. Using three-dimensional models it is shown that modelling discrete holes at the outlet as opposed to a continuous slot, which is the approximation inherent in the two-dimensional axisymmetric model, has little effect on the predicted Nusselt number distribution along the disc surface. Results of a conjugate heat transfer analysis of a spacer in the turbine section of a turboprop engine are then presented.

Advanced Prop-fan Engine Technology (APET) single- and counter-rotation gearbox/pitch change mechanism

NASA Technical Reports Server (NTRS)

Reynolds, C. N.

1985-01-01

The preliminary design of advanced technology (1992) turboprop engines for single-rotation prop-fans and conceptual designs of pitch change mechanisms for single- and counter-rotation prop-fan application are discussed. The single-rotation gearbox is a split path, in-line configuration. The counter-rotation gearbox is an in-line, differential planetary design. The pitch change mechanisms for both the single- and counter-rotation arrangements are rotary/hydraulic. The advanced technology single-rotation gearbox yields a 2.4 percent improvement in aircraft fuel burn and a one percent improvement in operating cost relative to a current technology gearbox. The 1992 counter-rotation gearbox is 15 percent lighter, 15 percent more reliable, 5 percent lower in cost, and 45 percent lower in maintenance cost than the 1992 single-rotation gearbox. The pitch controls are modular, accessible, and external.

Converting conformational changes to electrostatic energy in molecular motors: The energetics of ATP synthase.

PubMed

Strajbl, Marek; Shurki, Avital; Warshel, Arieh

2003-12-09

F1-ATPase is the catalytic component of the ATP synthase molecular machine responsible for most of the uphill synthesis of ATP in living systems. The enormous advances in biochemical and structural studies of this machine provide an opportunity for detailed understanding of the nature of its rotary mechanism. However, further quantitative progress in this direction requires development of reliable ways of translating the observed structural changes to the corresponding energies. This requirement is particularly challenging because we are dealing with a large system that couples major structural changes with a chemical process. The present work provides such a structure-function correlation by using the linear response approximation to describe the rotary mechanism. This approach allows one to evaluate the energy of transitions between different conformational states by considering only the changes in the corresponding electrostatic energies of the ligands. The relevant energetics are also obtained by calculating the linear response approximation-based free energies of transferring the ligands from water to the different sites of F1-ATPase in their different conformational states. We also use the empirical valence bond approach to evaluate the actual free-energy profile for the ATP synthesis in the different conformational states of the system. Integrating the information from the different approaches provides a semiquantitative structure-function correlation for F1-ATPase. It is found that the conformational changes are converted to changes in the electrostatic interaction between the protein and its ligands, which drives the ATP synthesis.

Advanced System Design Requirements for Large and Small Fixed-wing Aerial Application Systems for Agriculture

NASA Technical Reports Server (NTRS)

Hinely, J. T., Jr.; Boyles, R. Q., Jr.

1979-01-01

Several candidate aircraft configurations were defined over the range of 1000 to 10,000 pounds payload and evaluated over a broad spectrum of agricultural missions. From these studies, baseline design points were selected at 3200 pounds payload for the small aircraft and 7500 pounds for the large aircraft. The small baseline aircraft utilizes a single turboprop powerplant while the large aircraft utilizes two turboprop powerplants. These configurations were optimized for wing loading, aspect ratio, and power loading to provide the best mission economics in representative missions. Wing loading of 20 lb/sq ft was selected for the small aircraft and 25 lb/sq ft for the large aircraft. Aspect ratio of 8 was selected for both aircraft. It was found that a 10% reduction in engine power from the original configurations provided improved mission economics for both aircraft by reducing the cost of the turboprop. Refined configurations incorporate a 675 HP engine in the small aircraft and two 688 HP engines in the large aircraft.

ATP11B mediates platinum resistance in ovarian cancer

PubMed Central

Moreno-Smith, Myrthala; Halder, J.B.; Meltzer, Paul S.; Gonda, Tamas A.; Mangala, Lingegowda S.; Rupaimoole, Rajesha; Lu, Chunhua; Nagaraja, Archana S.; Gharpure, Kshipra M.; Kang, Yu; Rodriguez-Aguayo, Cristian; Vivas-Mejia, Pablo E.; Zand, Behrouz; Schmandt, Rosemarie; Wang, Hua; Langley, Robert R.; Jennings, Nicholas B.; Ivan, Cristina; Coffin, Jeremy E.; Armaiz, Guillermo N.; Bottsford-Miller, Justin; Kim, Sang Bae; Halleck, Margaret S.; Hendrix, Mary J.C.; Bornman, William; Bar-Eli, Menashe; Lee, Ju-Seog; Siddik, Zahid H.; Lopez-Berestein, Gabriel; Sood, Anil K.

2013-01-01

Platinum compounds display clinical activity against a wide variety of solid tumors; however, resistance to these agents is a major limitation in cancer therapy. Reduced platinum uptake and increased platinum export are examples of resistance mechanisms that limit the extent of DNA damage. Here, we report the discovery and characterization of the role of ATP11B, a P-type ATPase membrane protein, in cisplatin resistance. We found that ATP11B expression was correlated with higher tumor grade in human ovarian cancer samples and with cisplatin resistance in human ovarian cancer cell lines. ATP11B gene silencing restored the sensitivity of ovarian cancer cell lines to cisplatin in vitro. Combined therapy of cisplatin and ATP11B-targeted siRNA significantly decreased cancer growth in mice bearing ovarian tumors derived from cisplatin-sensitive and -resistant cells. In vitro mechanistic studies on cellular platinum content and cisplatin efflux kinetics indicated that ATP11B enhances the export of cisplatin from cells. The colocalization of ATP11B with fluorescent cisplatin and with vesicular trafficking proteins, such as syntaxin-6 (STX6) and vesicular-associated membrane protein 4 (VAMP4), strongly suggests that ATP11B contributes to secretory vesicular transport of cisplatin from Golgi to plasma membrane. In conclusion, inhibition of ATP11B expression could serve as a therapeutic strategy to overcome cisplatin resistance. PMID:23585472

Modeling the effects of hypoxia on ATP turnover in exercising muscle

NASA Technical Reports Server (NTRS)

Arthur, P. G.; Hogan, M. C.; Bebout, D. E.; Wagner, P. D.; Hochachka, P. W.

1992-01-01

Most models of metabolic control concentrate on the regulation of ATP production and largely ignore the regulation of ATP demand. We describe a model, based on the results of Hogan et al. (J. Appl. Physiol. 73: 728-736, 1992), that incorporates the effects of ATP demand. The model is developed from the premise that a unique set of intracellular conditions can be measured at each level of ATP turnover and that this relationship is best described by energetic state. Current concepts suggest that cells are capable of maintaining oxygen consumption in the face of declines in the concentration of oxygen through compensatory changes in cellular metabolites. We show that these compensatory changes can cause significant declines in ATP demand and result in a decline in oxygen consumption and ATP turnover. Furthermore we find that hypoxia does not directly affect the rate of anaerobic ATP synthesis and associated lactate production. Rather, lactate production appears to be related to energetic state, whatever the PO2. The model is used to describe the interaction between ATP demand and ATP supply in determining final ATP turnover.

7 CFR 3300.88 - Fees for U.S. ATP certificates.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 15 2014-01-01 2014-01-01 false Fees for U.S. ATP certificates. 3300.88 Section 3300... EQUIPMENT TO BE USED FOR SUCH CARRIAGE (ATP); INSPECTION, TESTING, AND CERTIFICATION OF SPECIAL EQUIPMENT Other Provisions § 3300.88 Fees for U.S. ATP certificates. The fee schedule for issuance of U.S. ATP...

7 CFR 3300.88 - Fees for U.S. ATP certificates.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 15 2012-01-01 2012-01-01 false Fees for U.S. ATP certificates. 3300.88 Section 3300... EQUIPMENT TO BE USED FOR SUCH CARRIAGE (ATP); INSPECTION, TESTING, AND CERTIFICATION OF SPECIAL EQUIPMENT Other Provisions § 3300.88 Fees for U.S. ATP certificates. The fee schedule for issuance of U.S. ATP...

7 CFR 3300.88 - Fees for U.S. ATP certificates.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 15 2013-01-01 2013-01-01 false Fees for U.S. ATP certificates. 3300.88 Section 3300... EQUIPMENT TO BE USED FOR SUCH CARRIAGE (ATP); INSPECTION, TESTING, AND CERTIFICATION OF SPECIAL EQUIPMENT Other Provisions § 3300.88 Fees for U.S. ATP certificates. The fee schedule for issuance of U.S. ATP...

7 CFR 3300.88 - Fees for U.S. ATP certificates.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 15 2010-01-01 2010-01-01 false Fees for U.S. ATP certificates. 3300.88 Section 3300... EQUIPMENT TO BE USED FOR SUCH CARRIAGE (ATP); INSPECTION, TESTING, AND CERTIFICATION OF SPECIAL EQUIPMENT Other Provisions § 3300.88 Fees for U.S. ATP certificates. The fee schedule for issuance of U.S. ATP...

7 CFR 3300.88 - Fees for U.S. ATP certificates.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 15 2011-01-01 2011-01-01 false Fees for U.S. ATP certificates. 3300.88 Section 3300... EQUIPMENT TO BE USED FOR SUCH CARRIAGE (ATP); INSPECTION, TESTING, AND CERTIFICATION OF SPECIAL EQUIPMENT Other Provisions § 3300.88 Fees for U.S. ATP certificates. The fee schedule for issuance of U.S. ATP...

Capture and quality control mechanisms for ATP binding

PubMed Central

Li, Li; Martinis, Susan A.

2013-01-01

The catalytic events in members of the nucleotidylyl transferase superfamily are initiated by a millisecond binding of ATP in the active site. Through metadynamics simulations on a class I aminoacyl-tRNA synthetase (aaRSs), the largest group in the superfamily, we calculate the free energy landscape of ATP selection and binding. Mutagenesis studies and fluorescence spectroscopy validated the identification of the most populated intermediate states. The rapid first binding step involves formation of encounter complexes captured through a fly-casting mechanism that acts up on the triphosphate moiety of ATP. In the slower nucleoside binding step, a conserved histidine in the HxxH motif orients the incoming ATP through base-stacking interactions resulting in a deep minimum in the free energy surface. Mutation of this histidine significantly decreases the binding affinity measured experimentally and computationally. The metadynamics simulations further reveal an intermediate quality control state that the synthetases and most likely other members of the superfamily use to select ATP over other nucleoside triphosphates. PMID:23276298

Imaging extracellular ATP with a genetically-encoded, ratiometric fluorescent sensor

PubMed Central

Conley, Jason M.

2017-01-01

Extracellular adenosine triphosphate (ATP) is a key purinergic signal that mediates cell-to-cell communication both within and between organ systems. We address the need for a robust and minimally invasive approach to measuring extracellular ATP by re-engineering the ATeam ATP sensor to be expressed on the cell surface. Using this approach, we image real-time changes in extracellular ATP levels with a sensor that is fully genetically-encoded and does not require an exogenous substrate. In addition, the sensor is ratiometric to allow for reliable quantitation of extracellular ATP fluxes. Using live-cell microscopy, we characterize sensor performance when expressed on cultured Neuro2A cells, and we measure both stimulated release of ATP and its clearance by ectonucleotidases. Thus, this proof-of-principle demonstrates a first-generation sensor to report extracellular ATP dynamics that may be useful for studying purinergic signaling in living specimens. PMID:29121644

Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H(+) pumping.

PubMed

Hotra, Adam; Suter, Manuel; Biuković, Goran; Ragunathan, Priya; Kundu, Subhashri; Dick, Thomas; Grüber, Gerhard

2016-05-01

The F1 FO -ATP synthase is one of the enzymes that is essential to meet the energy requirement of both the proliferating aerobic and hypoxic dormant stages of the life cycle of mycobacteria. Most F-ATP synthases consume ATP in the α3 :β3 headpiece to drive the γ subunit, which couples ATP cleavage with proton pumping in the c ring of FO via the bottom of the γ subunit. ATPase-driven H(+) pumping is latent in mycobacteria. The presence of a unique 14 amino acid residue loop of the mycobacterial γ subunit has been described and aligned in close vicinity to the c-ring loop Priya R et al. (2013) J Bioenerg Biomembr 45, 121-129 Here, we used inverted membrane vesicles (IMVs) of fast-growing Mycobacterium smegmatis and a variety of covalent and non-covalent inhibitors to characterize the ATP hydrolysis activity of the F-ATP synthase inside IMVs. These vesicles formed a platform to investigate the function of the unique mycobaterial γ loop by deleting the respective loop-encoding sequence (γ166-179 ) in the genome of M. smegmatis. ATP hydrolysis-driven H(+) pumping was observed in IMVs containing the Δγ166-179 mutant protein but not for IMVs containing the wild-type F-ATP synthase. In addition, when compared to the wild-type enzyme, IMVs containing the Δγ166-179 mutant protein showed increased ATP cleavage and lower levels of ATP synthesis, demonstrating that the loop affects ATPase activity, ATPase-driven H(+) pumping and ATP synthesis. These results further indicate that the loop may affect coupling of ATP hydrolysis and synthesis in a different mode. © 2016 Federation of European Biochemical Societies.

Effects of ultraviolet B irradiation, proinflammatory cytokines and raised extracellular calcium concentration on the expression of ATP2A2 and ATP2C1.

PubMed

Mayuzumi, N; Ikeda, S; Kawada, H; Fan, P S; Ogawa, H

2005-04-01

Darier disease (DD) and Hailey-Hailey disease (HHD) are autosomal dominantly inherited skin disorders that histologically share the characteristics of suprabasal separation and acantholysis of epidermal keratinocytes. Various mutations in the DD gene (ATP2A2) and the HHD gene (ATP2C1) (respectively encoding the calcium pumps of the sarco/endoplasmic reticulum and the Golgi apparatus) have recently been described in multiple families with DD and HHD. Mutations in ATP2A2 or ATP2C1 have been suggested as causing the conditions via the mechanism of haploinsufficiency. Ultraviolet (UV) B irradiation is thought to be an aggravating factor in both diseases. To examine the effects of various stimuli on ATP2A2 and ATP2C1 mRNA expression, and to examine the role of calcium pumps during keratinocyte differentiation. The effects of UVB irradiation, of UVB-inducible inflammatory cytokines produced by keratinocytes and of high-calcium medium (1.8 mmol L(-1) as opposed to 0.08 mmol L(-1) Ca2+) on ATP2A2 and ATP2C1 mRNA expression were quantified in cultured normal human keratinocytes using reverse transcription-polymerase chain reaction. Expression of ATP2A2 and ATP2C1 mRNA was suppressed immediately after exposure to UVB irradiation, and modulation of mRNA expression was achieved in keratinocytes cultured with proinflammatory cytokines. The mRNA expression of both genes was increased significantly after the shift to high extracellular Ca2+ concentration. The results suggest that modulation of ATP2A2 and ATP2C1 mRNA expression by UV or cytokines might contribute to the clinical presentations unique to DD and HHD, and that the controlled expression of these genes plays an important role in keratinocyte homeostasis, function and differentiation.

Air-Stimulated ATP Release from Keratinocytes Occurs through Connexin Hemichannels

PubMed Central

Barr, Travis P.; Albrecht, Phillip J.; Hou, Quanzhi; Mongin, Alexander A.; Strichartz, Gary R.; Rice, Frank L.

2013-01-01

Cutaneous ATP release plays an important role in both epidermal stratification and chronic pain, but little is known about ATP release mechanisms in keratinocytes that comprise the epidermis. In this study, we analyzed ATP release from cultured human neonatal keratinocytes briefly exposed to air, a process previously demonstrated to trigger ATP release from these cells. We show that exposing keratinocytes to air by removing media for 15 seconds causes a robust, long-lasting ATP release. This air-stimulated ATP release was increased in calcium differentiated cultures which showed a corresponding increase in connexin 43 mRNA, a major component of keratinocyte hemichannels. The known connexin hemichannel inhibitors 1-octanol and carbenoxolone both significantly reduced air-stimulated ATP release, as did two drugs traditionally used as ABC transporter inhibitors (glibenclamide and verapamil). These same 4 inhibitors also prevented an increase in the uptake of a connexin permeable dye induced by air exposure, confirming that connexin hemichannels are open during air-stimulated ATP release. In contrast, activity of the MDR1 ABC transporter was reduced by air exposure and the drugs that inhibited air-stimulated ATP release had differential effects on this transporter. These results indicate that air exposure elicits non-vesicular release of ATP from keratinocytes through connexin hemichannels and that drugs used to target connexin hemichannels and ABC transporters may cross-inhibit. Connexins represent a novel, peripheral target for the treatment of chronic pain and dermatological disease. PMID:23457608

Air-stimulated ATP release from keratinocytes occurs through connexin hemichannels.

PubMed

Barr, Travis P; Albrecht, Phillip J; Hou, Quanzhi; Mongin, Alexander A; Strichartz, Gary R; Rice, Frank L

2013-01-01

Cutaneous ATP release plays an important role in both epidermal stratification and chronic pain, but little is known about ATP release mechanisms in keratinocytes that comprise the epidermis. In this study, we analyzed ATP release from cultured human neonatal keratinocytes briefly exposed to air, a process previously demonstrated to trigger ATP release from these cells. We show that exposing keratinocytes to air by removing media for 15 seconds causes a robust, long-lasting ATP release. This air-stimulated ATP release was increased in calcium differentiated cultures which showed a corresponding increase in connexin 43 mRNA, a major component of keratinocyte hemichannels. The known connexin hemichannel inhibitors 1-octanol and carbenoxolone both significantly reduced air-stimulated ATP release, as did two drugs traditionally used as ABC transporter inhibitors (glibenclamide and verapamil). These same 4 inhibitors also prevented an increase in the uptake of a connexin permeable dye induced by air exposure, confirming that connexin hemichannels are open during air-stimulated ATP release. In contrast, activity of the MDR1 ABC transporter was reduced by air exposure and the drugs that inhibited air-stimulated ATP release had differential effects on this transporter. These results indicate that air exposure elicits non-vesicular release of ATP from keratinocytes through connexin hemichannels and that drugs used to target connexin hemichannels and ABC transporters may cross-inhibit. Connexins represent a novel, peripheral target for the treatment of chronic pain and dermatological disease.

Aircraft Energy Efficiency (ACEE) status report

NASA Technical Reports Server (NTRS)

Nored, D. L.; Dugan, J. F., Jr.; Saunders, N. T.; Ziemianski, J. A.

1979-01-01

Fuel efficiency in aeronautics, for fuel conservation in general as well as for its effect on commercial aircraft operating economics is considered. Projects of the Aircraft Energy Efficiency Program related to propulsion are emphasized. These include: (1) engine component improvement, directed at performance improvement and engine diagnostics for prolonged service life; (2) energy efficient engine, directed at proving the technology base for the next generation of turbofan engines; and (3) advanced turboprop, directed at advancing the technology of turboprop powered aircraft to a point suitable for commercial airline service. Progress in these technology areas is reported.

Effects of Tributyltin Chloride on Cybrids with or without an ATP Synthase Pathologic Mutation

PubMed Central

López-Gallardo, Ester; Llobet, Laura; Emperador, Sonia; Montoya, Julio; Ruiz-Pesini, Eduardo

2016-01-01

Background: The oxidative phosphorylation system (OXPHOS) includes nuclear chromosome (nDNA)– and mitochondrial DNA (mtDNA)–encoded polypeptides. Many rare OXPHOS disorders, such as striatal necrosis syndromes, are caused by genetic mutations. Despite important advances in sequencing procedures, causative mutations remain undetected in some patients. It is possible that etiologic factors, such as environmental toxins, are the cause of these cases. Indeed, the inhibition of a particular enzyme by a poison could imitate the biochemical effects of pathological mutations in that enzyme. Moreover, environmental factors can modify the penetrance or expressivity of pathological mutations. Objectives: We studied the interaction between mitochondrially encoded ATP synthase 6 (p.MT-ATP6) subunit and an environmental exposure that may contribute phenotypic differences between healthy individuals and patients suffering from striatal necrosis syndromes or other mitochondriopathies. Methods: We analyzed the effects of the ATP synthase inhibitor tributyltin chloride (TBTC), a widely distributed environmental factor that contaminates human food and water, on transmitochondrial cell lines with or without an ATP synthase mutation that causes striatal necrosis syndrome. Doses were selected based on TBTC concentrations previously reported in human whole blood samples. Results: TBTC modified the phenotypic effects caused by a pathological mtDNA mutation. Interestingly, wild-type cells treated with this xenobiotic showed similar bioenergetics when compared with the untreated mutated cells. Conclusions: In addition to the known genetic causes, our findings suggest that environmental exposure to TBTC might contribute to the etiology of striatal necrosis syndromes. Citation: López-Gallardo E, Llobet L, Emperador S, Montoya J, Ruiz-Pesini E. 2016. Effects of tributyltin chloride on cybrids with or without an ATP synthase pathologic mutation. Environ Health Perspect 124:1399–1405;�

Effects of Tributyltin Chloride on Cybrids with or without an ATP Synthase Pathologic Mutation.

PubMed

López-Gallardo, Ester; Llobet, Laura; Emperador, Sonia; Montoya, Julio; Ruiz-Pesini, Eduardo

2016-09-01

The oxidative phosphorylation system (OXPHOS) includes nuclear chromosome (nDNA)- and mitochondrial DNA (mtDNA)-encoded polypeptides. Many rare OXPHOS disorders, such as striatal necrosis syndromes, are caused by genetic mutations. Despite important advances in sequencing procedures, causative mutations remain undetected in some patients. It is possible that etiologic factors, such as environmental toxins, are the cause of these cases. Indeed, the inhibition of a particular enzyme by a poison could imitate the biochemical effects of pathological mutations in that enzyme. Moreover, environmental factors can modify the penetrance or expressivity of pathological mutations. We studied the interaction between mitochondrially encoded ATP synthase 6 (p.MT-ATP6) subunit and an environmental exposure that may contribute phenotypic differences between healthy individuals and patients suffering from striatal necrosis syndromes or other mitochondriopathies. We analyzed the effects of the ATP synthase inhibitor tributyltin chloride (TBTC), a widely distributed environmental factor that contaminates human food and water, on transmitochondrial cell lines with or without an ATP synthase mutation that causes striatal necrosis syndrome. Doses were selected based on TBTC concentrations previously reported in human whole blood samples. TBTC modified the phenotypic effects caused by a pathological mtDNA mutation. Interestingly, wild-type cells treated with this xenobiotic showed similar bioenergetics when compared with the untreated mutated cells. In addition to the known genetic causes, our findings suggest that environmental exposure to TBTC might contribute to the etiology of striatal necrosis syndromes. López-Gallardo E, Llobet L, Emperador S, Montoya J, Ruiz-Pesini E. 2016. Effects of tributyltin chloride on cybrids with or without an ATP synthase pathologic mutation. Environ Health Perspect 124:1399-1405; http://dx.doi.org/10.1289/EHP182.

Structural Characterization of Two Metastable ATP-Bound States of P-Glycoprotein

PubMed Central

O’Mara, Megan L.; Mark, Alan E.

2014-01-01

ATP Binding Cassette (ABC) transporters couple the binding and hydrolysis of ATP to the transport of substrate molecules across the membrane. The mechanism by which ATP binding and/or hydrolysis drives the conformational changes associated with substrate transport has not yet been characterized fully. Here, changes in the conformation of the ABC export protein P-glycoprotein on ATP binding are examined in a series of molecular dynamics simulations. When one molecule of ATP is placed at the ATP binding site associated with each of the two nucleotide binding domains (NBDs), the membrane-embedded P-glycoprotein crystal structure adopts two distinct metastable conformations. In one, each ATP molecule interacts primarily with the Walker A motif of the corresponding NBD. In the other, the ATP molecules interacts with both Walker A motif of one NBD and the Signature motif of the opposite NBD inducing the partial dimerization of the NBDs. This interaction is more extensive in one of the two ATP binding site, leading to an asymmetric structure. The overall conformation of the transmembrane domains is not altered in either of these metastable states, indicating that the conformational changes associated with ATP binding observed in the simulations in the absence of substrate do not lead to the outward-facing conformation and thus would be insufficient in themselves to drive transport. Nevertheless, the metastable intermediate ATP-bound conformations observed are compatible with a wide range of experimental cross-linking data demonstrating the simulations do capture physiologically important conformations. Analysis of the interaction between ATP and its cofactor Mg2+ with each NBD indicates that the coordination of ATP and Mg2+ differs between the two NBDs. The role structural asymmetry may play in ATP binding and hydrolysis is discussed. Furthermore, we demonstrate that our results are not heavily influenced by the crystal structure chosen for initiation of the simulations

Renal epithelial cells can release ATP by vesicular fusion

PubMed Central

Bjaelde, Randi G.; Arnadottir, Sigrid S.; Overgaard, Morten T.; Leipziger, Jens; Praetorius, Helle A.

2013-01-01

Renal epithelial cells have the ability to release nucleotides as paracrine factors. In the intercalated cells of the collecting duct, ATP is released by connexin30 (cx30), which is selectively expressed in this cell type. However, ATP is released by virtually all renal epithelia and the aim of the present study was to identify possible alternative nucleotide release pathways in a renal epithelial cell model. We used MDCK (type1) cells to screen for various potential ATP release pathways. In these cells, inhibition of the vesicular H+-ATPases (bafilomycin) reduced both the spontaneous and hypotonically (80%)-induced nucleotide release. Interference with vesicular fusion using N-ethylamide markedly reduced the spontaneous nucleotide release, as did interference with trafficking from the endoplasmic reticulum to the Golgi apparatus (brefeldin A1) and vesicular transport (nocodazole). These findings were substantiated using a siRNA directed against SNAP-23, which significantly reduced spontaneous ATP release. Inhibition of pannexin and connexins did not affect the spontaneous ATP release in this cell type, which consists of ~90% principal cells. TIRF-microscopy of either fluorescently-labeled ATP (MANT-ATP) or quinacrine-loaded vesicles, revealed that spontaneous release of single vesicles could be promoted by either hypoosmolality (50%) or ionomycin. This vesicular release decreased the overall cellular fluorescence by 5.8 and 7.6% respectively. In summary, this study supports the notion that spontaneous and induced ATP release can occur via exocytosis in renal epithelial cells. PMID:24065923

Three-Dimensional Structures Reveal Multiple ADP/ATP Binding Modes

DOE Office of Scientific and Technical Information (OSTI.GOV)

C Simmons; C Magee; D Smith

The creation of synthetic enzymes with predefined functions represents a major challenge in future synthetic biology applications. Here, we describe six structures of de novo proteins that have been determined using protein crystallography to address how simple enzymes perform catalysis. Three structures are of a protein, DX, selected for its stability and ability to tightly bind ATP. Despite the addition of ATP to the crystallization conditions, the presence of a bound but distorted ATP was found only under excess ATP conditions, with ADP being present under equimolar conditions or when crystallized for a prolonged period of time. A bound ADPmore » cofactor was evident when Asp was substituted for Val at residue 65, but ATP in a linear configuration is present when Phe was substituted for Tyr at residue 43. These new structures complement previously determined structures of DX and the protein with the Phe 43 to Tyr substitution [Simmons, C. R., et al. (2009) ACS Chem. Biol. 4, 649-658] and together demonstrate the multiple ADP/ATP binding modes from which a model emerges in which the DX protein binds ATP in a configuration that represents a transitional state for the catalysis of ATP to ADP through a slow, metal-free reaction capable of multiple turnovers. This unusual observation suggests that design-free methods can be used to generate novel protein scaffolds that are tailor-made for catalysis.« less

Myosin Motors: The Chemical Restraints Imposed by ATP

NASA Astrophysics Data System (ADS)

Rayment, I.; Allingham, J.

Most molecular motors use ATP. This is not surprising since this is the universal currency of energy for most of life's processes. The question to be addressed in this chapter is how does the fundamental chemistry of ATP hydrolysis in- fluence the observed organization of linear molecular motors seen today with a focus on myosin. This chapter is written to ask in simple terms what can be gained by reconsidering the chemistry of ATP hydrolysis.

The chloroplast ATP synthase features the characteristic redox regulation machinery.

PubMed

Hisabori, Toru; Sunamura, Ei-Ichiro; Kim, Yusung; Konno, Hiroki

2013-11-20

Regulation of the activity of the chloroplast ATP synthase is largely accomplished by the chloroplast thioredoxin system, the main redox regulation system in chloroplasts, which is directly coupled to the photosynthetic reaction. We review the current understanding of the redox regulation system of the chloroplast ATP synthase. The thioredoxin-targeted portion of the ATP synthase consists of two cysteines located on the central axis subunit γ. The redox state of these two cysteines is under the influence of chloroplast thioredoxin, which directly controls rotation during catalysis by inducing a conformational change in this subunit. The molecular mechanism of redox regulation of the chloroplast ATP synthase has recently been determined. Regulation of the activity of the chloroplast ATP synthase is critical in driving efficiency into the ATP synthesis reaction in chloroplasts. The molecular architecture of the chloroplast ATP synthase, which confers redox regulatory properties requires further investigation, in light of the molecular structure of the enzyme complex as well as the physiological significance of the regulation system.

Experimental aerodynamic performance of advanced 40 deg-swept 10-blade propeller model at Mach 0.6 to 0.85

NASA Technical Reports Server (NTRS)

Mitchell, Glenn A.

1988-01-01

A propeller designated as SR-6, designed with 40 deg of sweep and 10 blades to cruise at Mach 0.8 at an altitude of 10.7 km (35,000 ft), was tested in the NASA Lewis Research Center's 8- by 6-Foot Wind Tunnel. This propeller was one of a series of advanced single rotation propeller models designed and tested as part of the NASA Advanced Turboprop Project. Design-point net efficiency was almost constant to Mach 0.75 but fell above this speed more rapidly than that of any previously tested advanced propeller. Alternative spinners that further reduced the near-hub interblade Mach numbers and relieved the observed hub choking improved performance above Mach 0.75. One spinner attained estimated SR-6 Design-point net deficiencies of 80.6 percent at Mach 0.75 and 79.2 percent at Mach 0.8, higher than the measured performance of any previously tested advanced single-rotation propeller at these speeds.

Neurokinin B potentiates ATP-activated currents in rat DRG neurons.

PubMed

Wang, M J; Xiong, S H; Li, Z W

2001-12-27

This study aimed to explore whether NKB could modulate the responses mediated by ATP receptor (P2X purinoceptor). Whole-cell patch clamp and repatch experiments were performed on cultured rat DRG neurons. The majority of neurons examined were sensitive both to ATP and to NKB (77.1%, 54/70). NKB preapplied could potentiate ATP-activated currents (I(ATP)) markedly; this effect was concentration-dependent and could be blocked by SR 142801, an NK3 receptor antagonist. Preapplication of 0.001, 0.01, 0.1 and 1.0 microM NKB increased ATP-activated currents by 55.1+/-18.8, 75.2+/-17.4, 84.1+/-18.8 and 81.0+/-21.7%, respectively. The concentration-response curves for ATP with and without preapplication of NKB show that: (1) preapplication of NKB shifted the curve upwards; (2) the maximal amplitude of I(ATP) with NKB preapplication increased by 78.5%, while the threshold value remained unchanged; (3) the EC(50) values of the two curves were very close (44 vs. 42 microM). Intracellular dialysis of H-7 by using repatch clamp technique could block the potentiation of I(ATP) by NKB. It suggests that this potentiating effect was caused by phosphorylation of ATP receptor, which resulted from the activation of G protein coupled NK3 receptor and consequential intracellular signal transduction cascade.

Extracellular ATP Acts on Jasmonate Signaling to Reinforce Plant Defense.

PubMed

Tripathi, Diwaker; Zhang, Tong; Koo, Abraham J; Stacey, Gary; Tanaka, Kiwamu

2018-01-01

Damaged cells send various signals to stimulate defense responses. Recent identification and genetic studies of the plant purinoceptor, P2K1 (also known as DORN1), have demonstrated that extracellular ATP is a signal involved in plant stress responses, including wounding, perhaps to evoke plant defense. However, it remains largely unknown how extracellular ATP induces plant defense responses. Here, we demonstrate that extracellular ATP induces plant defense mediated through activation of the intracellular signaling of jasmonate (JA), a well-characterized defense hormone. In Arabidopsis ( Arabidopsis thaliana ) leaves, ATP pretreatment induced resistance against the necrotrophic fungus, Botrytis cinerea The induced resistance was enhanced in the P2K1 receptor overexpression line, but reduced in the receptor mutant, dorn1 - 3 Mining the transcriptome data revealed that ATP induces a set of JA-induced genes. In addition, the P2K1-associated coexpression network contains defense-related genes, including those encoding jasmonate ZIM-domain (JAZ) proteins, which play key roles as repressors of JA signaling. We examined whether extracellular ATP impacts the stability of JAZ1 in Arabidopsis. The results showed that the JAZ1 stability decreased in response to ATP addition in a proteasome-dependent manner. This reduction required intracellular signaling via second messengers-cytosolic calcium, reactive oxygen species, and nitric oxide. Interestingly, the ATP-induced JAZ1 degradation was attenuated in the JA receptor mutant, coi1 , but not in the JA biosynthesis mutant, aos , or upon addition of JA biosynthesis inhibitors. Immunoprecipitation analysis demonstrated that ATP increases the interaction between COI1 and JAZ1, suggesting direct cross talk between extracellular ATP and JA in intracellular signaling events. Taken together, these results suggest that extracellular ATP signaling directly impacts the JA signaling pathway to maximize plant defense responses. © 2018

Behavior and stability of adenosine triphosphate (ATP) during chlorine disinfection.

PubMed

Nescerecka, Alina; Juhna, Talis; Hammes, Frederik

2016-09-15

Adenosine triphosphate (ATP) analysis is a cultivation-independent alternative method for the determination of bacterial viability in both chlorinated and non-chlorinated water. Here we investigated the behavior and stability of ATP during chlorination in detail. Different sodium hypochlorite doses (0-22.4 mg-Cl2 L(-1); 5 min exposure) were applied to an Escherichia coli pure culture suspended in filtered river water. We observed decreasing intracellular ATP with increasing chlorine concentrations, but extracellular ATP concentrations only increased when the chlorine dose exceeded 0.35 mg L(-1). The release of ATP from chlorine-damaged bacteria coincided with severe membrane damage detected with flow cytometry (FCM). The stability of extracellular ATP was subsequently studied in different water matrixes, and we found that extracellular ATP was stable in sterile deionized water and also in chlorinated water until extremely high chlorine doses (≤11.2 mg-Cl2 L(-1); 5 min exposure). In contrast, ATP decreased relatively slowly (k = 0.145 h(-1)) in 0.1 μm filtered river water, presumably due to degradation by either extracellular enzymes or the fraction of bacteria that were able to pass through the filter. Extracellular ATP decreased considerably faster (k = 0.368 h(-1)) during batch growth of a river water bacterial community. A series of growth potential tests showed that extracellular ATP molecules were utilized as a phosphorus source during bacteria proliferation. From the combined data we conclude that ATP released from bacteria at high chlorine doses could promote bacteria regrowth, contributing to biological instability in drinking water distribution systems. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inhibition of ATP Hydrolysis by Thermoalkaliphilic F1Fo-ATP Synthase Is Controlled by the C Terminus of the ɛ Subunit

PubMed Central

Keis, Stefanie; Stocker, Achim; Dimroth, Peter; Cook, Gregory M.

2006-01-01

The F1Fo-ATP synthases of alkaliphilic bacteria exhibit latent ATPase activity, and for the thermoalkaliphile Bacillus sp. strain TA2.A1, this activity is intrinsic to the F1 moiety. To study the mechanism of ATPase inhibition, we developed a heterologous expression system in Escherichia coli to produce TA2F1 complexes from this thermoalkaliphile. Like the native F1Fo-ATP synthase, the recombinant TA2F1 was blocked in ATP hydrolysis activity, and this activity was stimulated by the detergent lauryldimethylamine oxide. To determine if the C-terminal domain of the ɛ subunit acts as an inhibitor of ATPase activity and if an electrostatic interaction plays a role, a TA2F1 mutant with either a truncated ɛ subunit [i.e., TA2F1(ɛΔC)] or substitution of basic residues in the second α-helix of ɛ with nonpolar alanines [i.e., TA2F1(ɛ6A)] was constructed. Both mutants showed ATP hydrolysis activity at low and high concentrations of ATP. Treatment of the purified F1Fo-ATP synthase and TA2F1(ɛWT) complex with proteases revealed that the ɛ subunit was resistant to proteolytic digestion. In contrast, the ɛ subunit of TA2F1(ɛ6A) was completely degraded by trypsin, indicating that the C-terminal arm was in a conformation where it was no longer protected from proteolytic digestion. In addition, ATPase activity was not further activated by protease treatment when compared to the untreated control, supporting the observation that ɛ was responsible for inhibition of ATPase activity. To study the effect of the alanine substitutions in the ɛ subunit in the entire holoenzyme, we reconstituted recombinant TA2F1 complexes with F1-stripped native membranes of strain TA2.A1. The reconstituted TA2FoF1(ɛWT) was blocked in ATP hydrolysis and exhibited low levels of ATP-driven proton pumping consistent with the F1Fo-ATP synthase in native membranes. Reconstituted TA2FoF1(ɛ6A) exhibited ATPase activity that correlated with increased ATP-driven proton pumping, confirming that the �

ATP7B detoxifies silver in ciliated airway epithelial cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ibricevic, Aida, E-mail: aidaibricevic@hotmail.co; Brody, Steven L., E-mail: sbrody@dom.wustl.ed; Youngs, Wiley J., E-mail: youngs@uakron.ed

2010-03-15

Silver is a centuries-old antibiotic agent currently used to treat infected burns. The sensitivity of a wide range of drug-resistant microorganisms to silver killing suggests that it may be useful for treating refractory lung infections. Toward this goal, we previously developed a methylated caffeine silver acetate compound, SCC1, that exhibits broad-spectrum antimicrobial activity against clinical strains of bacteria in vitro and when nebulized to lungs in mouse infection models. Preclinical testing of high concentrations of SCC1 in primary culture mouse tracheal epithelial cells (mTEC) showed selective ciliated cell death. Ciliated cell death was induced by both silver- and copper-containing compoundsmore » but not by the methylated caffeine portion of SCC1. We hypothesized that copper transporting P-type ATPases, ATP7A and ATP7B, play a role in silver detoxification in the airway. In mTEC, ATP7A was expressed in non-ciliated cells, whereas ATP7B was expressed only in ciliated cells. The exposure of mTEC to SCC1 induced the trafficking of ATP7B, but not ATP7A, suggesting the presence of a cell-specific silver uptake and detoxification mechanisms. Indeed, the expression of the copper uptake protein CTR1 was also restricted to ciliated cells. A role of ATP7B in silver detoxification was further substantiated when treatment of SCC1 significantly increased cell death in ATP7B shRNA-treated HepG2 cells. In addition, mTEC from ATP7B{sup -/-} mice showed enhanced loss of ciliated cells compared to wild type. These studies are the first to demonstrate a cell type-specific expression of the Ag{sup +}/Cu{sup +} transporters ATP7A, ATP7B, and CTR1 in airway epithelial cells and a role for ATP7B in detoxification of these metals in the lung.« less

ATP-induced changes in rat skeletal muscle contractility.

PubMed

Gabdrakhmanov, A I; Khayrullin, A E; Grishin, C H; Ziganshin, A U

2015-01-01

Extracellular purine compounds, adenosine triphosphate (ATP) and adenosine, are involved in regulation of many cell functions, engaging in rapid and long-term cellular processes. The nucleotides, including ATP, exert their extracellular effects by influencing membrane P2 receptors. ATP outside of the cell rapidly is metabolized by the ecto-enzyme system to produce adenosine, which acts on separate adenosine (P1) receptors. Since adenosine and ATP often are functional antagonists, ATP degradation not only limits its effect, but also brings new ligand with different, often opposing, properties. Great variety and widespread of P2 and adenosine receptors in the body emphasize the important physiological and pathophysiological significance of these receptors, and make them very attractive as targets for potential drug action.The existence of several subtypes of P2 and adenosine receptors has been shown in the skeletal muscles. ATP as a co-transmitter is densely packed together with classical neurotransmitters in the presynaptic vesicles of vertebral motor units but until recently ATP was refused to have its own functional role there and was recognized only as a source of adenosine. However, on the eve of the third millennium there appeared data that ATP, released from the nerve ending and acting on presynaptic P2 receptors, suppresses subsequent quantum release of acetylcholine. The final product of its degradation, adenosine, performs a similar inhibitory effect acting on presynaptic adenosine receptors.Despite the fact that the mechanisms of presynaptic inhibitory action of ATP and other purines were studied earlier, the object of those studies was usually neuromuscular synapse of cold-blooded animals. The few studies, in which experiments were carried out on preparations of warm-blooded animals, described the basic effects of purines. These often were guided by the convenience of preparation of the synapses of the diaphragm. We think that those results cannot be

Functional analysis of mutations in the ATP loop of the Wilson disease copper transporter, ATP7B.

PubMed

Luoma, Leiah M; Deeb, Taha M M; Macintyre, Georgina; Cox, Diane W

2010-05-01

Wilson disease (WND) is an autosomal recessive disorder resulting from mutation of ATP7B. Transport of copper by ATP7B from the trans-Golgi of hepatocytes into apical membrane-trafficked vesicles for excretion in the bile is the major means of copper elimination from the body. Although copper is an essential nutrient, homeostasis must be carefully maintained. If homeostasis is disrupted, copper can accumulate within the liver, kidney, cornea, and/or brain. The range of organs affected leads to clinical heterogeneity and difficulty in WND diagnosis. Sequencing of ATP7B is an important adjunct for diagnosis but has led to the discovery of many novel missense variants. Although prediction programs are available, functional characterization is essential for determining the consequence of novel variants. We have tested 12 missense variants localized to the ATP loop of ATP7B and compared three predictive programs (SIFT, PolyPhen, and Align-GVGD). We found p.L1043P, p.G1000R, p.G1101R, p.I1102T, p.V1239G, and p.D1267V deleterious; p.G1176E and p.G1287S intermediate; p.E1173G temperature sensitive; p.T991M and p.I1148T mild; and p.R1228T functioning as wild type. We found that SIFT most often agreed with functional data (92%), compared with PolyPhen (83%) and Align-GVGD (67%). We conclude that variants found to negatively affect function likely contribute to the WND phenotype in patients. (c) 2010 Wiley-Liss, Inc.

Fast transient currents in Na,K-ATPase induced by ATP concentration jumps from the P3-[1-(3',5'-dimethoxyphenyl)-2-phenyl-2-oxo]ethyl ester of ATP.

PubMed Central

Sokolov, V S; Apell, H J; Corrie, J E; Trentham, D R

1998-01-01

Electrogenic ion transport by Na,K-ATPase was investigated by analysis of transient currents in a model system of protein-containing membrane fragments adsorbed to planar lipid bilayers. Sodium transport was triggered by ATP concentration jumps in which ATP was released from an inactive precursor by an intense near-UV light flash. The method has been used previously with the P3-1-(2-nitrophenyl)ethyl ester of ATP (NPE-caged ATP), from which the relatively slow rate of ATP release limits analysis of processes in the pump mechanism controlled by rate constants greater than 100 s(-1) at physiological pH. Here Na,K-ATPase was reinvestigated using the P3-[1-(3,5-dimethoxyphenyl)-2-phenyl-2-oxo]ethyl ester of ATP (DMB-caged ATP), which has an ATP release rate of >10(5) s(-1). Under otherwise identical conditions, photorelease of ATP from DMB-caged ATP showed faster kinetics of the transient current compared to that from NPE-caged ATP. With DMB-caged ATP, transient currents had rate profiles that were relatively insensitive to pH and the concentration of caged compound. Rate constants of ATP binding and of the E1 to E2 conformational change were compatible with earlier studies. Rate constants of enzyme phosphorylation and ADP-dependent dephosphorylation were 600 s(-1) and 1.5 x 10(6) M(-1) s(-1), respectively, at pH 7.2 and 22 degrees C. PMID:9591656

A Therapeutic Connection between Dietary Phytochemicals and ATP Synthase.

PubMed

Ahmad, Zulfiqar; Hassan, Sherif S; Azim, Sofiya

2017-11-20

For centuries, phytochemicals have been used to prevent and cure multiple health ailments. Phytochemicals have been reported to have antioxidant, antidiabetic, antitussive, antiparasitic, anticancer, and antimicrobial properties. Generally, the therapeutic use of phytochemicals is based on tradition or word of mouth with few evidence-based studies. Moreover, molecular level interactions or molecular targets for the majority of phytochemicals are unknown. In recent years, antibiotic resistance by microbes has become a major healthcare concern. As such, the use of phytochemicals with antimicrobial properties has become pertinent. Natural compounds from plants, vegetables, herbs, and spices with strong antimicrobial properties present an excellent opportunity for preventing and combating antibiotic resistant microbial infections. ATP synthase is the fundamental means of cellular energy. Inhibition of ATP synthase may deprive cells of required energy leading to cell death, and a variety of dietary phytochemicals are known to inhibit ATP synthase. Structural modifications of phytochemicals have been shown to increase the inhibitory potency and extent of inhibition. Sitedirected mutagenic analysis has elucidated the binding site(s) for some phytochemicals on ATP synthase. Amino acid variations in and around the phytochemical binding sites can result in selective binding and inhibition of microbial ATP synthase. In this review, the therapeutic connection between dietary phytochemicals and ATP synthase is summarized based on the inhibition of ATP synthase by dietary phytochemicals. Research suggests selective targeting of ATP synthase is a valuable alternative molecular level approach to combat antibiotic resistant microbial infections. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

A Therapeutic Connection between Dietary Phytochemicals and ATP Synthase

PubMed Central

Ahmad, Zulfiqar; Hassan, Sherif S.; Azim, Sofiya

2017-01-01

For centuries, phytochemicals have been used to prevent and cure multiple health ailments. Phytochemicals have been reported to have antioxidant, antidiabetic, antitussive, antiparasitic, anticancer, and antimicrobial properties. Generally, the therapeutic use of phy-tochemicals is based on tradition or word of mouth with few evidence-based studies. Moreo-ver, molecular level interactions or molecular targets for the majority of phytochemicals are unknown. In recent years, antibiotic resistance by microbes has become a major healthcare concern. As such, the use of phytochemicals with antimicrobial properties has become perti-nent. Natural compounds from plants, vegetables, herbs, and spices with strong antimicrobial properties present an excellent opportunity for preventing and combating antibiotic resistant microbial infections. ATP synthase is the fundamental means of cellular energy. Inhibition of ATP synthase may deprive cells of required energy leading to cell death, and a variety of die-tary phytochemicals are known to inhibit ATP synthase. Structural modifications of phyto-chemicals have been shown to increase the inhibitory potency and extent of inhibition. Site-directed mutagenic analysis has elucidated the binding site(s) for some phytochemicals on ATP synthase. Amino acid variations in and around the phytochemical binding sites can re-sult in selective binding and inhibition of microbial ATP synthase. In this review, the therapeu-tic connection between dietary phytochemicals and ATP synthase is summarized based on the inhibition of ATP synthase by dietary phytochemicals. Research suggests selective target-ing of ATP synthase is a valuable alternative molecular level approach to combat antibiotic resistant microbial infections. PMID:28831918

Modulation of postjunctional α-adrenergic vasoconstriction during exercise and exogenous ATP infusions in ageing humans

PubMed Central

Kirby, Brett S; Crecelius, Anne R; Voyles, Wyatt F; Dinenno, Frank A

2011-01-01

exercise is impaired with age, whereas the sympatholytic effect of exogenous ATP is preserved. Thus, if impairments in vascular control during exercise in older adults involve vasoactive ATP, we speculate that circulating ATP is reduced with advancing age. PMID:21486772

Functional studies of ATP sulfurylase from Penicillium chrysogenum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seubert, P.A.

1985-01-01

ATP sulfurylase from Penicillium chrysogenum has a specific activity (V/sub max/) of 6-7 units x mg protein/sup -1/ determined with the physiological substrates of MgATP and SO/sub 4//sup 2 -/ and assayed by (A) initial velocity measurements with APS kinase and inorganic pyrophosphatase present and (B) analysis of nonlinear reaction progress curves. The fact both assays give the same results show the intrinsic activity of ATP sulfurylase is much higher than previously reported. In initial velocity dead-end inhibition studies, the sulfate analog S/sub 2/O/sub 3//sup 2 -/ is a competitive inhibitor of SO/sub 42/..sqrt.. and a noncompetitive inhibitor of MgATP.more » Monovalent oxyanions such as NO/sub 3//sup -/, ClO/sub 3//sup -/, ClO/sub 4//sup -/, and FSO/sub 3//sup -/ behave as uncompetitive inhibitors of MgATP and thus seem not to be true sulfate analogs. The reverse reaction was assayed by the pyrophosphate dependent release of /sup 35/SO/sub 4//sup 2 -/ from AP/sup 35/S. Product inhibition by MgATP or SO/sub 4//sup 2 -/ is competitive with APS and mixed-type with PP/sub i/. Imidodiphosphate can serve as an alternative substrate for PP/sub i/. ATP sulfurylase binds (but does not hydrolyze) APS. A Scatchard plot of the APS binding is nonlinear, suggesting at least two types of sites. The cumulative results are qualitatively consistent with the random addition of MgATP and SO/sub 4//sup 2 -/ and the ordered release of first MgPP/sub i/ then APS, with APS release being partially rate limiting. Certain quantitative discrepancies suggest either an unknown variable (e.g. enzyme concentration) complicates the analysis or, in light of binding studies that the actual mechanism is more complicated (e.g. alternating sites) than any of the conventional models examined.« less

Fuel conservation merits of advanced turboprop transport aircraft. Final report, January--August 1977

DOE Office of Scientific and Technical Information (OSTI.GOV)

Revell, J.D.; Tullis, R.H.

1977-08-01

The advantages of a propfan powered aircraft for the commercial air transportation system were assessed by the comparison with an equivalent turbofan transport. Comparisons were accomplished on the basis of fuel utilization and operating costs, as well as aircraft weight and size. Advantages of the propfan aircraft, concerning fuel utilization and operating costs, were accomplished by considering: (1) incorporation of propfan performance and acoustic data; (2) revised mission profiles (longer design range and reduction in; and cruise speed) (3) utilization of alternate and advanced technology engines.

Adenosine triphosphate (ATP) as a possible indicator of extraterrestrial biology

NASA Technical Reports Server (NTRS)

Chappelle, E. W.; Picciolo, G. L.

1974-01-01

The ubiquity of adenosine triphosphate (ATP) in terrestrial organisms provides the basis for proposing the assay of this vital metabolic intermediate for detecting extraterrestrial biological activity. If an organic carbon chemistry is present on the planets, the occurrence of ATP is possible either from biosynthetic or purely chemical reactions. However, ATP's relative complexity minimizes the probability of abiogenic synthesis. A sensitive technique for the quantitative detection of ATP was developed using the firefly bioluminescent reaction. The procedure was used successfully for the determination of the ATP content of soil and bacteria. This technique is also being investigated from the standpoint of its application in clinical medicine.

Fo-driven Rotation in the ATP Synthase Direction against the Force of F1 ATPase in the FoF1 ATP Synthase*

PubMed Central

Martin, James; Hudson, Jennifer; Hornung, Tassilo; Frasch, Wayne D.

2015-01-01

Living organisms rely on the FoF1 ATP synthase to maintain the non-equilibrium chemical gradient of ATP to ADP and phosphate that provides the primary energy source for cellular processes. How the Fo motor uses a transmembrane electrochemical ion gradient to create clockwise torque that overcomes F1 ATPase-driven counterclockwise torque at high ATP is a major unresolved question. Using single FoF1 molecules embedded in lipid bilayer nanodiscs, we now report the observation of Fo-dependent rotation of the c10 ring in the ATP synthase (clockwise) direction against the counterclockwise force of ATPase-driven rotation that occurs upon formation of a leash with Fo stator subunit a. Mutational studies indicate that the leash is important for ATP synthase activity and support a mechanism in which residues aGlu-196 and cArg-50 participate in the cytoplasmic proton half-channel to promote leash formation. PMID:25713065

Extracellular ATP inhibits root gravitropism at concentrations that inhibit polar auxin transport

NASA Technical Reports Server (NTRS)

Tang, Wenqiang; Brady, Shari R.; Sun, Yu; Muday, Gloria K.; Roux, Stanley J.

2003-01-01

Raising the level of extracellular ATP to mM concentrations similar to those found inside cells can block gravitropism of Arabidopsis roots. When plants are grown in Murashige and Skoog medium supplied with 1 mM ATP, their roots grow horizontally instead of growing straight down. Medium with 2 mM ATP induces root curling, and 3 mM ATP stimulates lateral root growth. When plants are transferred to medium containing exogenous ATP, the gravity response is reduced or in some cases completely blocked by ATP. Equivalent concentrations of ADP or inorganic phosphate have slight but usually statistically insignificant effects, suggesting the specificity of ATP in these responses. The ATP effects may be attributable to the disturbance of auxin distribution in roots by exogenously applied ATP, because extracellular ATP can alter the pattern of auxin-induced gene expression in DR5-beta-glucuronidase transgenic plants and increase the response sensitivity of plant roots to exogenously added auxin. The presence of extracellular ATP also decreases basipetal auxin transport in a dose-dependent fashion in both maize (Zea mays) and Arabidopsis roots and increases the retention of [(3)H]indole-3-acetic acid in root tips of maize. Taken together, these results suggest that the inhibitory effects of extracellular ATP on auxin distribution may happen at the level of auxin export. The potential role of the trans-plasma membrane ATP gradient in auxin export and plant root gravitropism is discussed.

Serotonergic regulation of distention-induced ATP release from the urothelium.

PubMed

Matsumoto-Miyai, Kazumasa; Yamada, Erika; Shinzawa, Eriko; Koyama, Yoshihisa; Shimada, Shoichi; Yoshizumi, Masaru; Kawatani, Masahito

2016-04-01

Serotonin [5-hydroxytryptamine (5-HT)] is involved in both motor and sensory functions in hollow organs, especially in the gastrointestinal tract. However, the involvement of 5-HT in visceral sensation of the urinary bladder remains unknown. Because distention-induced ATP release from the urothelium plays an essential role in visceral sensation of the urinary bladder, we investigated the regulation of urothelial ATP release by the 5-HT signaling system. RT-PCR and immunohistochemical analyses of the urothelium revealed specific expression of 5-HT 1D and 5-HT 4 receptors. The addition of 5-HT did not affect urothelial ATP release without bladder distention, but it significantly reduced distention-induced ATP release by physiological pressure during urine storage (5 cmH 2 O). The inhibitory effect of 5-HT on distention-elicited ATP release was blocked by preincubation with the 5-HT 1B/1D antagonist GR-127935 but not by the 5-HT 4 antagonist SB-204070. mRNA encoding tryptophan hydroxylase 1 was detected in the urinary bladder by nested RT-PCR amplification, and l-tryptophan or the selective serotonin reuptake inhibitor citalopram also inhibited ATP release, indicating that 5-HT is endogenously synthesized and released in the urinary bladder. The addition of GR-127935 significantly enhanced the distention-elicited ATP release 40 min after distention, whereas SB-204070 reduced the amount of ATP release 20 min after distention. These data suggest that 5-HT 4 facilitates the distention-induced ATP release at an earlier stage, whereas 5-HT 1D inhibits ATP release at a later stage. The net inhibitory effect of 5-HT indicates that the action of 5-HT on the urothelium is mediated predominantly by 5-HT 1D . Copyright © 2016 the American Physiological Society.

Adenosine triphosphate (ATP) reduces amyloid-β protein misfolding in vitro.

PubMed

Coskuner, Orkid; Murray, Ian V J

2014-01-01

Alzheimer's disease (AD) is a devastating disease of aging that initiates decades prior to clinical manifestation and represents an impending epidemic. Two early features of AD are metabolic dysfunction and changes in amyloid-β protein (Aβ) levels. Since levels of ATP decrease over the course of the disease and Aβ is an early biomarker of AD, we sought to uncover novel linkages between the two. First and remarkably, a GxxxG motif is common between both Aβ (oligomerization motif) and nucleotide binding proteins (Rossmann fold). Second, ATP was demonstrated to protect against Aβ mediated cytotoxicity. Last, there is structural similarity between ATP and amyloid binding/inhibitory compounds such as ThioT, melatonin, and indoles. Thus, we investigated whether ATP alters misfolding of the pathologically relevant Aβ42. To test this hypothesis, we performed computational and biochemical studies. Our computational studies demonstrate that ATP interacts strongly with Tyr10 and Ser26 of Aβ fibrils in solution. Experimentally, both ATP and ADP reduced Aβ misfolding at physiological intracellular concentrations, with thresholds at ~500 μM and 1 mM respectively. This inhibition of Aβ misfolding is specific; requiring Tyr10 of Aβ and is enhanced by magnesium. Last, cerebrospinal fluid ATP levels are in the nanomolar range and decreased with AD pathology. This initial and novel finding regarding the ATP interaction with Aβ and reduction of Aβ misfolding has potential significance to the AD field. It provides an underlying mechanism for published links between metabolic dysfunction and AD. It also suggests a potential role of ATP in AD pathology, as the occurrence of misfolded extracellular Aβ mirrors lowered extracellular ATP levels. Last, the findings suggest that Aβ conformation change may be a sensor of metabolic dysfunction.

Loss of the gene for the alpha subunit of ATP synthase (ATP5A1) from the W chromosome in the African grey parrot (Psittacus erithacus).

PubMed

de Kloet, S R

2001-08-01

This study describes the results of an analysis using Southern blotting, the polymerase chain reaction, and sequencing which shows that the African grey parrot (Psittacus erithacus) lacks the W-chromosomal gene for the alpha subunit of mitochondrial ATP synthase (ATP5A1W). Additional evidence shows that in other psittacines a fragment of the ATP5A1W gene contains five times as many nonsynonymous nucleotide replacements as the homologous fragment of the Z gene. Therefore, whereas in these other psittacines the corresponding ATP5A1Z protein fragment is highly conserved and varies by only a few, moderately conservative amino acid substitutions, the homologous ATP5A1W fragments contain a considerable number of, sometimes highly nonconservative, amino acid replacements. In one of these species, the ringneck parakeet (Psittacula krameri), the ATP5A1W gene is present in an inactive form because of the presence of a nonsense codon. Other changes, possibly leading to an inactive ATP5A1W gene product, involve the substitution of arginine residues by cysteine in the ATP5A1W protein of the mitred conure (Aratinga mitrata) and the blue and gold macaw (Ara ararauna). The data suggest also that although the divergence of the psittacine ATP5A1W and ATP5A1Z genes preceded the origin of the psittacidae, this divergence occurred independently of a similar process in the myna (Gracula religiosa), the outgroup used in this study.

Aerodynamic analysis of three advanced configurations using the TranAir full-potential code

NASA Technical Reports Server (NTRS)

Madson, M. D.; Carmichael, R. L.; Mendoza, J. P.

1989-01-01

Computational results are presented for three advanced configurations: the F-16A with wing tip missiles and under wing fuel tanks, the Oblique Wing Research Aircraft, and an Advanced Turboprop research model. These results were generated by the latest version of the TranAir full potential code, which solves for transonic flow over complex configurations. TranAir embeds a surface paneled geometry definition in a uniform rectangular flow field grid, thus avoiding the use of surface conforming grids, and decoupling the grid generation process from the definition of the configuration. The new version of the code locally refines the uniform grid near the surface of the geometry, based on local panel size and/or user input. This method distributes the flow field grid points much more efficiently than the previous version of the code, which solved for a grid that was uniform everywhere in the flow field. TranAir results are presented for the three configurations and are compared with wind tunnel data.

The Chloroplast atpA Gene Cluster in Chlamydomonas reinhardtii1

PubMed Central

Drapier, Dominique; Suzuki, Hideki; Levy, Haim; Rimbault, Blandine; Kindle, Karen L.; Stern, David B.; Wollman, Francis-André

1998-01-01

Most chloroplast genes in vascular plants are organized into polycistronic transcription units, which generate a complex pattern of mono-, di-, and polycistronic transcripts. In contrast, most Chlamydomonas reinhardtii chloroplast transcripts characterized to date have been monocistronic. This paper describes the atpA gene cluster in the C. reinhardtii chloroplast genome, which includes the atpA, psbI, cemA, and atpH genes, encoding the α-subunit of the coupling-factor-1 (CF1) ATP synthase, a small photosystem II polypeptide, a chloroplast envelope membrane protein, and subunit III of the CF0 ATP synthase, respectively. We show that promoters precede the atpA, psbI, and atpH genes, but not the cemA gene, and that cemA mRNA is present only as part of di-, tri-, or tetracistronic transcripts. Deletions introduced into the gene cluster reveal, first, that CF1-α can be translated from di- or polycistronic transcripts, and, second, that substantial reductions in mRNA quantity have minimal effects on protein synthesis rates. We suggest that posttranscriptional mRNA processing is common in C. reinhardtii chloroplasts, permitting the expression of multiple genes from a single promoter. PMID:9625716

ATP and microfilaments in cellular oxidant injury.

PubMed Central

Hinshaw, D. B.; Armstrong, B. C.; Burger, J. M.; Beals, T. F.; Hyslop, P. A.

1988-01-01

Oxidant injury produces dramatic changes in cytoskeletal organization and cell shape. ATP synthetic pathways are major targets of oxidant injury resulting in rapid depletion of cellular ATP following oxidant exposure. The relation of ATP depletion to the changes in microfilament organization seen following H2O2 exposure were examined in the P388D1 cell line. Three hours of glucose depletion alone resulted in a decline in cellular ATP levels to less than 10% of controls, which was comparable to ATP levels in cells 30 to 60 minutes after exposure to 5 mM H2O2 in the presence of glucose. Adherent cells stained with rhodamine phalloidin, a probe specific for polymerized (F) actin, revealed a progressive shortening of microfilaments into globular aggregates within cells depleted of glucose over 3 hours, a pattern similar to earlier observations of H2O2-injured cells after 1 hour. The changes in cellular ATP associated with glucose depletion or H2O2 exposure were then correlated with G actin content measured by the DNAse 1 inhibition assay. No real differences in G actin content as a percentage of total actin were seen in P388D1 cells following 3 hours of glucose depletion or 30 to 60 minutes after exposure to 5 mM H2O2. But 2 to 3 hours after exposure to H2O2 there was a progressive decrease in G actin as a percentage of total actin within the cells. Transmission electron microscopy of cells depleted of glucose for 3 h or 1 hour after exposure to H2O2 revealed the presence of side-to-side aggregates or bundles of microfilaments within the cells. These observations suggest that declining levels of ATP either from metabolic inhibition or H2O2 injury are correlated with the fragmentation and shortening of microfilaments into aggregates. No net change in monomeric or polymeric actin was necessary for this to occur. However, at later time points after H2O2 exposure some actin assembly did occur. Images p[484]-a p481-a p482-a Figure 2 Figure 3 PMID:3414780

Diadenosine tetraphosphate-gating of recombinant pancreatic ATP-sensitive K(+) channels.

PubMed

Jovanovic, S; Jovanovic, A

2001-02-01

Diadenosine tetraphosphate (Ap4A) has been recently discovered in the pancreatic beta cells where targets ATP-sensitive K(+) (K(ATP)) channels, depolarizes the cell membrane and induces insulin secretion. However, whether Ap4A inhibit pancreatic K(ATP) channels by targeting protein channel complex itself was unknown. Therefore, we coexpressed pancreatic K(ATP) channel subunits, Kir6.2 and SUR1, in COS-7 cells and examined the effect of Ap4A on the single channel behavior using the inside-out configuration of the patch-clamp technique. Ap4A inhibited channel opening in a concentration-dependent manner. Analysis of single channels demonstrated that Ap4A did not change intraburst kinetic behavior of K(ATP) channels, but rather decreased burst duration and increased between-burst duration. It is concluded that Ap4A antagonizes K(ATP) channel opening by targeting channel subunits themselves and by keeping channels longer in closed interburst states.

Monitoring ATP dynamics in electrically active white matter tracts

PubMed Central

Trevisiol, Andrea; Saab, Aiman S; Winkler, Ulrike; Marx, Grit; Imamura, Hiromi; Möbius, Wiebke; Kusch, Kathrin; Nave, Klaus-Armin; Hirrlinger, Johannes

2017-01-01

In several neurodegenerative diseases and myelin disorders, the degeneration profiles of myelinated axons are compatible with underlying energy deficits. However, it is presently impossible to measure selectively axonal ATP levels in the electrically active nervous system. We combined transgenic expression of an ATP-sensor in neurons of mice with confocal FRET imaging and electrophysiological recordings of acutely isolated optic nerves. This allowed us to monitor dynamic changes and activity-dependent axonal ATP homeostasis at the cellular level and in real time. We find that changes in ATP levels correlate well with compound action potentials. However, this correlation is disrupted when metabolism of lactate is inhibited, suggesting that axonal glycolysis products are not sufficient to maintain mitochondrial energy metabolism of electrically active axons. The combined monitoring of cellular ATP and electrical activity is a novel tool to study neuronal and glial energy metabolism in normal physiology and in models of neurodegenerative disorders. DOI: http://dx.doi.org/10.7554/eLife.24241.001 PMID:28414271

The regulation of ATP release from the urothelium by adenosine and transepithelial potential.

PubMed

Dunning-Davies, Bryony M; Fry, Christopher H; Mansour, Dina; Ferguson, Douglas R

2013-03-01

WHAT'S KNOWN ON THE SUBJECT? AND WHAT DOES THE STUDY ADD?: Stretch of the urothelium, as occurs during bladder filling, is associated with a release of ATP that is postulated to act as a sensory neurotransmitter. The regulation of ATP release is poorly understood and in particular if there is a feedback mechanism provided by ATP itself. Adenosine, a breakdown product of ATP, is a potent inhibitor of stretch-induced ATP release, acting through and A1 receptor; endogenous levels are about 0.6μM. Data are consistent with ATP release relying on the rise of intracellular Ca2+. Transepithelial potential also controls ATP release, also acting via an A1 receptor-dependent pathway. To test the hypothesis that distension-induced ATP release from the bladder urothelium is regulated by adenosine as well as changes to transurothelial potential (TEP). To examine the role of changes to intracellular [Ca(2+) ] in ATP release. Rabbit urothelium/suburothelium membranes were used in an Ussing chamber system. Distension was induced by fluid removal from the chamber bathing the serosal (basolateral) membrane face. The TEP and short-circuit current were measured. ATP was measured in samples aspirated from the serosal chamber by a luciferin-luciferase assay. Intracellular [Ca(2+) ] was measured in isolated urothelial cells using the fluorochrome Fura-2. All experiments were performed at 37°C. Distension-induced ATP release was decreased by adenosine (1-10 μm) and enhanced by adenosine deaminase and A1- (but not A2-) receptor antagonists. Distension-induced ATP release was reduced by 2-APB, nifedipine and capsazepine; capsaicin induced ATP release in the absence of distension. ATP and capsaicin, but not adenosine, generated intracellular Ca(2+) transients; adenosine did not affect the ATP-generated Ca(2+) transient. ATP release was dependent on a finite transepithelial potential. Changes to TEP, in the absence of distension, generated ATP release that was in turn reduced by adenosine

Turboprop aircraft performance response to various environmental conditions

NASA Astrophysics Data System (ADS)

Ashenden, Russell Allen

1997-10-01

. The results suggest that the most severe icing is actually caused by drizzle sized drops as opposed to freezing rain. Furthermore, these results are similar to many twin-turboprop aircraft typically utilized by the commuter fleet.

Structural Basis for a Unique ATP Synthase Core Complex from Nanoarcheaum equitans*

PubMed Central

Mohanty, Soumya; Jobichen, Chacko; Chichili, Vishnu Priyanka Reddy; Velázquez-Campoy, Adrián; Low, Boon Chuan; Hogue, Christopher W. V.; Sivaraman, J.

2015-01-01

ATP synthesis is a critical and universal life process carried out by ATP synthases. Whereas eukaryotic and prokaryotic ATP synthases are well characterized, archaeal ATP synthases are relatively poorly understood. The hyperthermophilic archaeal parasite, Nanoarcheaum equitans, lacks several subunits of the ATP synthase and is suspected to be energetically dependent on its host, Ignicoccus hospitalis. This suggests that this ATP synthase might be a rudimentary machine. Here, we report the crystal structures and biophysical studies of the regulatory subunit, NeqB, the apo-NeqAB, and NeqAB in complex with nucleotides, ADP, and adenylyl-imidodiphosphate (non-hydrolysable analog of ATP). NeqB is ∼20 amino acids shorter at its C terminus than its homologs, but this does not impede its binding with NeqA to form the complex. The heterodimeric NeqAB complex assumes a closed, rigid conformation irrespective of nucleotide binding; this differs from its homologs, which require conformational changes for catalytic activity. Thus, although N. equitans possesses an ATP synthase core A3B3 hexameric complex, it might not function as a bona fide ATP synthase. PMID:26370083

TRPC5-eNOS Axis Negatively Regulates ATP-Induced Cardiomyocyte Hypertrophy.

PubMed

Sunggip, Caroline; Shimoda, Kakeru; Oda, Sayaka; Tanaka, Tomohiro; Nishiyama, Kazuhiro; Mangmool, Supachoke; Nishimura, Akiyuki; Numaga-Tomita, Takuro; Nishida, Motohiro

2018-01-01

Cardiac hypertrophy, induced by neurohumoral factors, including angiotensin II and endothelin-1, is a major predisposing factor for heart failure. These ligands can induce hypertrophic growth of neonatal rat cardiomyocytes (NRCMs) mainly through Ca 2+ -dependent calcineurin/nuclear factor of activated T cell (NFAT) signaling pathways activated by diacylglycerol-activated transient receptor potential canonical 3 and 6 (TRPC3/6) heteromultimer channels. Although extracellular nucleotide, adenosine 5'-triphosphate (ATP), is also known as most potent Ca 2+ -mobilizing ligand that acts on purinergic receptors, ATP never induces cardiomyocyte hypertrophy. Here we show that ATP-induced production of nitric oxide (NO) negatively regulates hypertrophic signaling mediated by TRPC3/6 channels in NRCMs. Pharmacological inhibition of NO synthase (NOS) potentiated ATP-induced increases in NFAT activity, protein synthesis, and transcriptional activity of brain natriuretic peptide. ATP significantly increased NO production and protein kinase G (PKG) activity compared to angiotensin II and endothelin-1. We found that ATP-induced Ca 2+ signaling requires inositol 1,4,5-trisphosphate (IP 3 ) receptor activation. Interestingly, inhibition of TRPC5, but not TRPC6 attenuated ATP-induced activation of Ca 2+ /NFAT-dependent signaling. As inhibition of TRPC5 attenuates ATP-stimulated NOS activation, these results suggest that NO-cGMP-PKG axis activated by IP 3 -mediated TRPC5 channels underlies negative regulation of TRPC3/6-dependent hypertrophic signaling induced by ATP stimulation.

Effects of oral ATP supplementation on anaerobic power and muscular strength.

PubMed

Jordan, Alexander N; Jurca, Radim; Abraham, Edward H; Salikhova, Anna; Mann, Julia K; Morss, Gina M; Church, Timothy S; Lucia, Alejandro; Earnest, Conrad P

2004-06-01

We examined 14 d of oral adenosine 5'-triphosphate (ATP) supplementation on indices of anaerobic capacity and muscular strength. Twenty-seven healthy males successfully completed the trial, after randomly receiving in a double-blind manner an oral dose of low dose (150 mg) or high dose (225 mg) ATP, or matched placebo. To improve absorption characteristics, the ATP was enterically coated. Total blood ATP (whole blood and plasma ATP) concentrations, two Wingate anaerobic power tests (30 s), and muscular strength (1RM and three sets of repetitions to fatigue at 70% of 1RM) were measured under three conditions: (i) baseline; (ii) acutely (7d later, no prior supplementation and 75 min after ATP ingestion); and (iii) after 14 d of daily ingestion (post). Statistical analyses showed no significant between or within group treatment effects for whole blood ATP or plasma ATP concentrations for any treatment condition. We also did not observe any treatment effects for any Wingate testing parameter including peak PO, total work, average PO for 30 s, or post-Wingate lactate accumulation. Overall, we observed no significant between group treatment effects for any muscular strength parameter. We did observe several within group differences for the group ingesting the high ATP dosage including 1RM (6.6%; P < 0.04) and repetitions to fatigue during set 1 of posttesting (18.5%; P < 0.007) and total lifting volume at post (22%; P < 0.003). We conclude that enterically coated oral ATP supplementation may provide small ergogenic effects on muscular strength under some treatment conditions.

S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics.

PubMed

Módis, Katalin; Ju, YoungJun; Ahmad, Akbar; Untereiner, Ashley A; Altaany, Zaid; Wu, Lingyun; Szabo, Csaba; Wang, Rui

2016-11-01

Mammalian cells can utilize hydrogen sulfide (H 2 S) to support mitochondrial respiration. The aim of our study was to explore the potential role of S-sulfhydration (a H 2 S-induced posttranslational modification, also known as S-persulfidation) of the mitochondrial inner membrane protein ATP synthase (F1F0 ATP synthase/Complex V) in the regulation of mitochondrial bioenergetics. Using a biotin switch assay, we have detected S-sulfhydration of the α subunit (ATP5A1) of ATP synthase in response to exposure to H 2 S in vitro. The H 2 S generator compound NaHS induced S-sulfhydration of ATP5A1 in HepG2 and HEK293 cell lysates in a concentration-dependent manner (50-300μM). The activity of immunocaptured mitochondrial ATP synthase enzyme isolated from HepG2 and HEK293 cells was stimulated by NaHS at low concentrations (10-100nM). Site-directed mutagenesis of ATP5A1 in HEK293 cells demonstrated that cysteine residues at positions 244 and 294 are subject to S-sulfhydration. The double mutant ATP synthase protein (C244S/C294S) showed a significantly reduced enzyme activity compared to control and the single-cysteine-mutated recombinant proteins (C244S or C294S). To determine whether endogenous H 2 S plays a role in the basal S-sulfhydration of ATP synthase in vivo, we compared liver tissues harvested from wild-type mice and mice deficient in cystathionine-gamma-lyase (CSE, one of the three principal mammalian H 2 S-producing enzymes). Significantly reduced S-sulfhydration of ATP5A1 was observed in liver homogenates of CSE -/- mice, compared to wild-type mice, suggesting a physiological role for CSE-derived endogenous H 2 S production in the S-sulfhydration of ATP synthase. Various forms of critical illness (including burn injury) upregulate H 2 S-producing enzymes and stimulate H 2 S biosynthesis. In liver tissues collected from mice subjected to burn injury, we detected an increased S-sulfhydration of ATP5A1 at the early time points post-burn. At later time points

Interior noise prediction methodology: ATDAC theory and validation

NASA Technical Reports Server (NTRS)

Mathur, Gopal P.; Gardner, Bryce K.

1992-01-01

The Acoustical Theory for Design of Aircraft Cabins (ATDAC) is a computer program developed to predict interior noise levels inside aircraft and to evaluate the effects of different aircraft configurations on the aircraft acoustical environment. The primary motivation for development of this program is the special interior noise problems associated with advanced turboprop (ATP) aircraft where there is a tonal, low frequency noise problem. Prediction of interior noise levels requires knowledge of the energy sources, the transmission paths, and the relationship between the energy variable and the sound pressure level. The energy sources include engine noise, both airborne and structure-borne; turbulent boundary layer noise; and interior noise sources such as air conditioner noise and auxiliary power unit noise. Since propeller and engine noise prediction programs are widely available, they are not included in ATDAC. Airborne engine noise from any prediction or measurement may be input to this program. This report describes the theory and equations implemented in the ATDAC program.

Interior noise prediction methodology: ATDAC theory and validation

NASA Astrophysics Data System (ADS)

Mathur, Gopal P.; Gardner, Bryce K.

1992-04-01

The Acoustical Theory for Design of Aircraft Cabins (ATDAC) is a computer program developed to predict interior noise levels inside aircraft and to evaluate the effects of different aircraft configurations on the aircraft acoustical environment. The primary motivation for development of this program is the special interior noise problems associated with advanced turboprop (ATP) aircraft where there is a tonal, low frequency noise problem. Prediction of interior noise levels requires knowledge of the energy sources, the transmission paths, and the relationship between the energy variable and the sound pressure level. The energy sources include engine noise, both airborne and structure-borne; turbulent boundary layer noise; and interior noise sources such as air conditioner noise and auxiliary power unit noise. Since propeller and engine noise prediction programs are widely available, they are not included in ATDAC. Airborne engine noise from any prediction or measurement may be input to this program. This report describes the theory and equations implemented in the ATDAC program.

Hydrostatic pressure activates ATP-sensitive K+ channels in lung epithelium by ATP release through pannexin and connexin hemichannels.

PubMed

Richter, Katrin; Kiefer, Kevin P; Grzesik, Benno A; Clauss, Wolfgang G; Fronius, Martin

2014-01-01

Lungs of air-breathing vertebrates are constantly exposed to mechanical forces and therefore are suitable for investigation of mechanotransduction processes in nonexcitable cells and tissues. Freshly dissected Xenopus laevis lungs were used for transepithelial short-circuit current (ISC) recordings and were exposed to increased hydrostatic pressure (HP; 5 cm fluid column, modified Ussing chamber). I(SC) values obtained under HP (I(5cm)) were normalized to values before HP (I(0cm)) application (I(5cm)/I(0cm)). Under control conditions, HP decreased I(SC) (I(5cm)/I(0cm)=0.84; n=68; P<0.0001). This effect was reversible and repeatable ≥30 times. Preincubation with ATP-sensitive K(+) channel (K(ATP)) inhibitors (HMR1098 and glibenclamide) prevented the decrease in I(SC) (I(5cm)/I(0cm): HMR1098=1.19, P<0.0001; glibenclamide=1.11, P<0.0001). Similar effects were observed with hemichannel inhibitors (I(5cm)/I(0cm): meclofenamic acid=1.09, P<0.0001; probenecid=1.0, P<0.0001). The HP effect was accompanied by release of ATP (P<0.05), determined by luciferin-luciferase luminescence in perfusion solution from the luminal side of an Ussing chamber. ATP release was abrogated by both meclofenamic acid and probenecid. RT-PCR experiments revealed the expression of pannexin and connexin hemichannels and KATP subunit transcripts in X. laevis lung. These data show an activation of KATP in pulmonary epithelial cells in response to HP that is induced by ATP release through mechanosensitive pannexin and connexin hemichannels. These findings represent a novel mechanism of mechanotransduction in nonexcitable cells.

Energy transduction in the F1 motor of ATP synthase.

PubMed

Wang, H; Oster, G

1998-11-19

ATP synthase is the universal enzyme that manufactures ATP from ADP and phosphate by using the energy derived from a transmembrane protonmotive gradient. It can also reverse itself and hydrolyse ATP to pump protons against an electrochemical gradient. ATP synthase carries out both its synthetic and hydrolytic cycles by a rotary mechanism. This has been confirmed in the direction of hydrolysis after isolation of the soluble F1 portion of the protein and visualization of the actual rotation of the central 'shaft' of the enzyme with respect to the rest of the molecule, making ATP synthase the world's smallest rotary engine. Here we present a model for this engine that accounts for its mechanochemical behaviour in both the hydrolysing and synthesizing directions. We conclude that the F1 motor achieves its high mechanical torque and almost 100% efficiency because it converts the free energy of ATP binding into elastic strain, which is then released by a coordinated kinetic and tightly coupled conformational mechanism to create a rotary torque.

Energy transduction in the F1 motor of ATP synthase

NASA Astrophysics Data System (ADS)

Wang, Hongyun; Oster, George

1998-11-01

ATP synthase is the universal enzyme that manufactures ATP from ADP and phosphate by using the energy derived from a transmembrane protonmotive gradient. It can also reverse itself and hydrolyse ATP to pump protons against an electrochemical gradient. ATP synthase carries out both its synthetic and hydrolytic cycles by a rotary mechanism. This has been confirmed in the direction of hydrolysis, after isolation of the soluble F1 portion of the protein and visualization of the actual rotation of the central `shaft' of the enzyme with respect to the rest of the molecule, making ATP synthase the world's smallest rotary engine. Here we present a model for this engine that accounts for its mechanochemical behaviour in both the hydrolysing and synthesizing directions. We conclude that the F1 motor achieves its high mechanical torque and almost 100% efficiency because it converts the free energy of ATP binding into elastic strain, which is then released by a coordinated kinetic and tightly coupled conformational mechanism to create a rotary torque.

Application of the principle of linked functions to ATP-driven ion pumps: kinetics of activation by ATP.

PubMed Central

Reynolds, J A; Johnson, E A; Tanford, C

1985-01-01

If a ligand binds with unequal affinity to two distinct states of a protein, then the equilibrium between the two states becomes a function of the concentration of the ligand. A necessary consequence is that the ligand must also affect the forward and/or reverse rate constants for transition between the two states. For an enzyme or transport protein with such a transition as a slow step in the catalytic cycle, the overall rate also becomes a function of ligand concentration. These conclusions are independent of whether or not the ligand is a direct participant in the reaction. If it is a direct participant, then the kinetic effect arising from the principle of linked functions is distinct from the direct catalytic effect. These principles suffice to account for the biphasic response of the hydrolytic activity of ATP-driven ion pumps to the concentration of ATP, without the need to invoke more than one ATP binding site per catalytic center. PMID:2987939

Paul D. Boyer, Adenosine Triphosphate (ATP), and the Binding Change

Science.gov Websites

-- October 1975, DOE Technical Report, 1975 A Perspective of the Binding Change Mechanism for ATP Synthesis Reports, Vol. 18, No. 3, 1998 ATP Synthesis and the Binding Change Mechanism: The Work of Paul D. Boyer Mechanism of ATP Synthesis Additional Web Pages: Adenosine Triphosphate: The Energy Currency of Life Paul D

Updated Assessment of an Open Rotor Airplane Using an Advanced Blade Design

NASA Technical Reports Server (NTRS)

Hendricks, Eric S.; Berton, Jeffrey J.; Haller, William J.; Tong, Michael T.; Guynn, Mark D.

2013-01-01

Application of open rotor propulsion systems (historically referred to as "advanced turboprops" or "propfans") to subsonic transport aircraft received significant attention and research in the 1970s and 1980s when fuel efficiency was the driving focus of aeronautical research. Recent volatility in fuel prices and concern for aviation's environmental impact have renewed interest in open rotor propulsion, and revived research by NASA and a number of engine manufacturers. Over the last few years, NASA has revived and developed analysis capabilities to assess aircraft designs with open rotor propulsion systems. These efforts have been described in several previous papers along with initial results from applying these capabilities. The initial results indicated that open rotor engines have the potential to provide large reductions in fuel consumption and emissions. Initial noise analysis indicated that current noise regulations can be met with modern baseline blade designs. Improved blades incorporating low-noise features are expected to result in even lower noise levels. This paper describes improvements to the initial assessment, plus a follow-on study using a more advanced open rotor blade design to power the advanced singleaisle transport. The predicted performance and environmental results of these two advanced open rotor concepts are presented and compared.

The role of the urothelium and ATP in mediating detrusor smooth muscle contractility.

PubMed

Santoso, Aneira Gracia Hidayat; Sonarno, Ika Ariyani Bte; Arsad, Noor Aishah Bte; Liang, Willmann

2010-11-01

To examine the contractility of urothelium-intact (+UE) and urothelium-denuded (-UE) rat detrusor strips under adenosine triphosphate (ATP) treatment. Purinergic signaling exists in the bladder but both the inhibitory effect of ATP on detrusor contractions and the function of urothelial ATP are not established. Detrusor strips were obtained from bladders of young adult rats. Isometric tension from both transverse and longitudinal contractions was measured using a myograph. The muscarinic agonist carbachol (CCh) was used to induce contractions, which were under the influences of different concentrations of ATP. In both +UE and -UE strips, 1 mM ATP suppressed CCh-induced contractions. In longitudinal contractions, ATP added to the inhibitory effect of urothelium on CCh responses. Removal of the urothelium, but with exogenous ATP added, recovered the CCh responses to the same level as in +UE strips with no added ATP. Transverse contractions were less susceptible to ATP in the presence of urothelium. We showed that the urothelium and ATP suppressed CCh-induced contractions to a similar extent. The findings suggest an inhibitory role of urothelial ATP in mediating detrusor smooth muscle contractility, which may be impaired in diseased bladders. Copyright © 2010 Elsevier Inc. All rights reserved.

CFTR mediates noradrenaline-induced ATP efflux from DRG neurons.

PubMed

Kanno, Takeshi; Nishizaki, Tomoyuki

2011-09-24

In our earlier study, noradrenaline (NA) stimulated ATP release from dorsal root ganglion (DRG) neurons as mediated via β(3) adrenoceptors linked to G(s) protein involving protein kinase A (PKA) activation, to cause allodynia. The present study was conducted to understand how ATP is released from DRG neurons. In an outside-out patch-clamp configuration from acutely dissociated rat DRG neurons, single-channel currents, sensitive to the P2X receptor inhibitor PPADS, were evoked by approaching the patch-electrode tip close to a neuron, indicating that ATP is released from DRG neurons, to activate P2X receptor. NA increased the frequency of the single-channel events, but such NA effect was not found for DRG neurons transfected with the siRNA to silence the cystic fibrosis transmembrane conductance regulator (CFTR) gene. In the immunocytochemical study using acutely dissociated rat DRG cells, CFTR was expressed in neurons alone, but not satellite cells, fibroblasts, or Schwann cells. It is concluded from these results that CFTR mediates NA-induced ATP efflux from DRG neurons as an ATP channel.

Cardiac Metabolism in Heart Failure - Implications beyond ATP production

PubMed Central

Doenst, Torsten; Nguyen, T. Dung; Abel, E. Dale

2013-01-01

The heart has a high rate of ATP production and turnover which is required to maintain its continuous mechanical work. Perturbations in ATP generating processes may therefore affect contractile function directly. Characterizing cardiac metabolism in heart failure revealed several metabolic alterations termed metabolic remodeling, ranging from changes in substrate utilization to mitochondrial dysfunction, ultimately resulting in ATP deficiency and impaired contractility. However, ATP depletion is not the only relevant consequence of metabolic remodeling during heart failure. By providing cellular building blocks and signaling molecules, metabolic pathways control essential processes such as cell growth and regeneration. Thus, alterations in cardiac metabolism may also affect the progression to heart failure by mechanisms beyond ATP supply. Our aim is therefore to highlight that metabolic remodeling in heart failure not only results in impaired cardiac energetics, but also induces other processes implicated in the development of heart failure such as structural remodeling and oxidative stress. Accordingly, modulating cardiac metabolism in heart failure may have significant therapeutic relevance that goes beyond the energetic aspect. PMID:23989714

Luminescent Immunoprecipitation System (LIPS) for Detection of Autoantibodies Against ATP4A and ATP4B Subunits of Gastric Proton Pump H+,K+-ATPase in Atrophic Body Gastritis Patients

PubMed Central

Lahner, Edith; Brigatti, Cristina; Marzinotto, Ilaria; Carabotti, Marilia; Scalese, Giulia; Davidson, Howard W; Wenzlau, Janet M; Bosi, Emanuele; Piemonti, Lorenzo; Annibale, Bruno; Lampasona, Vito

2017-01-01

Objectives: Circulating autoantibodies targeting the H+/K+-ATPase proton pump of gastric parietal cells are considered markers of autoimmune gastritis, whose diagnostic accuracy in atrophic body gastritis, the pathological lesion of autoimmune gastritis, remains unknown. This study aimed to assess autoantibodies against ATP4A and ATP4B subunits of parietal cells H+, K+-ATPase in atrophic body gastritis patients and controls. Methods: One-hundred and four cases with atrophic body gastritis and 205 controls were assessed for serological autoantibodies specific for ATP4A or ATP4B subunits using luminescent immunoprecipitation system (LIPS). Recombinant luciferase-reporter-fused-antigens were expressed by in vitro transcription-translation (ATP4A) or after transfection in Expi293F cells (ATP4B), incubated with test sera, and immune complexes recovered using protein-A-sepharose. LIPS assays were compared with a commercial enzyme immunoassay (EIA) for parietal cell autoantibodies. Results: ATP4A and ATP4B autoantibody titers were higher in cases compared to controls (P<0.0001). The area under the receiver-operating characteristic curve was 0.98 (95% CI 0.965–0.996) for ATP4A, and 0.99 (95% CI 0.979–1.000) for ATP4B, both higher as compared with that of EIA: 0.86 (95% CI 0.809–0.896), P<0.0001. Sensitivity-specificity were 100–89% for ATP4A and 100–90% for ATP4B assay. Compared with LIPS, EIA for parietal cell autoantibodies showed a lower sensitivity (72%, P<0.0001) at a similar specificity (92%, P=0.558). Conclusions: Positivity to both, ATP4A and ATP4B autoantibodies is closely associated with atrophic body gastritis. Both assays had the highest sensitivity, at the cost of diagnostic accuracy (89 and 90% specificity), outperforming traditional EIA. Once validated, these LIPS assays should be valuable screening tools for detecting biomarkers of damaged atrophic oxyntic mucosa. PMID:28102858

Kinetic mechanism of ATP-sulphurylase from rat chondrosarcoma.

PubMed Central

Lyle, S; Geller, D H; Ng, K; Westley, J; Schwartz, N B

1994-01-01

ATP-sulphurylase catalyses the production of adenosine 5'-phosphosulphate (APS) from ATP and free sulphate with the release of PPi. APS kinase phosphorylates the APS intermediate to produce adenosine 3'-phosphate 5'-phosphosulphate (PAPS). The kinetic mechanism of rat chondrosarcoma ATP-sulphurylase was investigated by steady-state methods in the physiologically forward direction as well as the reverse direction. The sulphurylase activity was coupled to APS kinase activity in order to overcome the thermodynamic constraints of the sulphurylase reaction in the forward direction. Double-reciprocal initial-velocity plots for the forward sulphurylase intersect to the left of the ordinate for this reaction. KmATP and Kmsulphate were found to be 200 and 97 microM respectively. Chlorate, a competitive inhibitor with respect to sulphate, showed uncompetitive inhibition with respect to ATP with an apparent Ki of 1.97 mM. Steady-state data from experiments in the physiologically reverse direction also yielded double-reciprocal initial-velocity patterns that intersect to the left of the ordinate axis, with a KmAPS of 39 microM and a Kmpyrophosphate of 18 microM. The results of steady-state experiments in which Mg2+ was varied indicated that the true substrate is the MgPPi complex. An analogue of APS, adenosine 5'-[beta-methylene]phosphosulphate, was a linear inhibitor competitive with APS and non-competitive with respect to MgPPi. The simplest formal mechanism that agrees with all the data is an ordered steady-state single displacement with MgATP as the leading substrate in the forward direction and APS as the leading substrate in the reverse direction. PMID:8042976

Dynamics of shear-induced ATP release from red blood cells.

PubMed

Wan, Jiandi; Ristenpart, William D; Stone, Howard A

2008-10-28

Adenosine triphosphate (ATP) is a regulatory molecule for many cell functions, both for intracellular and, perhaps less well known, extracellular functions. An important example of the latter involves red blood cells (RBCs), which help regulate blood pressure by releasing ATP as a vasodilatory signaling molecule in response to the increased shear stress inside arterial constrictions. Although shear-induced ATP release has been observed widely and is believed to be triggered by deformation of the cell membrane, the underlying mechanosensing mechanism inside RBCs is still controversial. Here, we use an in vitro microfluidic approach to investigate the dynamics of shear-induced ATP release from human RBCs with millisecond resolution. We demonstrate that there is a sizable delay time between the onset of increased shear stress and the release of ATP. This response time decreases with shear stress, but surprisingly does not depend significantly on membrane rigidity. Furthermore, we show that even though the RBCs deform significantly in short constrictions (duration of increased stress <3 ms), no measurable ATP is released. This critical timescale is commensurate with a characteristic membrane relaxation time determined from observations of the cell deformation by using high-speed video. Taken together our results suggest a model wherein the retraction of the spectrin-actin cytoskeleton network triggers the mechanosensitive ATP release and a shear-dependent membrane viscosity controls the rate of release.

Stress granule formation via ATP depletion-triggered phase separation

NASA Astrophysics Data System (ADS)

Wurtz, Jean David; Lee, Chiu Fan

2018-04-01

Stress granules (SG) are droplets of proteins and RNA that form in the cell cytoplasm during stress conditions. We consider minimal models of stress granule formation based on the mechanism of phase separation regulated by ATP-driven chemical reactions. Motivated by experimental observations, we identify a minimal model of SG formation triggered by ATP depletion. Our analysis indicates that ATP is continuously hydrolysed to deter SG formation under normal conditions, and we provide specific predictions that can be tested experimentally.

Strain Background Modifies Phenotypes in the ATP8B1-Deficient Mouse

PubMed Central

Vargas, Julie C.; Xu, Hongmei; Groen, Annamiek; Paulusma, Coen C.; Grenert, James P.; Pawlikowska, Ludmila; Sen, Saunak; Elferink, Ronald P. J. Oude; Bull, Laura N.

2010-01-01

Background Mutations in ATP8B1 (FIC1) underlie cases of cholestatic disease, ranging from chronic and progressive (progressive familial intrahepatic cholestasis) to intermittent (benign recurrent intrahepatic cholestasis). The ATP8B1-deficient mouse serves as an animal model of human ATP8B1 deficiency. Methodology/Principal Findings We investigated the effect of genetic background on phenotypes of ATP8B1-deficient and wild-type mice, using C57Bl/6 (B6), 129, and (B6-129) F1 strain backgrounds. B6 background resulted in greater abnormalities in ATP8B1-deficient mice than did 129 and/or F1 background. ATP8B1-deficient pups of B6 background gained less weight. In adult ATP8B1-deficient mice at baseline, those of B6 background had lower serum cholesterol levels, higher serum alkaline phosphatase levels, and larger livers. After challenge with cholate-supplemented diet, these mice exhibited higher serum alkaline phosphatase and bilirubin levels, greater weight loss and larger livers. ATP8B1-deficient phenotypes in mice of F1 and 129 backgrounds are usually similar, suggesting that susceptibility to manifestations of ATP8B1 deficiency may be recessive. We also detected differences in hepatobiliary phenotypes between wild-type mice of differing strains. Conclusions/Significance Our results indicate that the ATP8B1-deficient mouse in a B6 background may be a better model of human ATP8B1 deficiency and highlight the importance of informed background strain selection for mouse models of liver disease. PMID:20126555

The Role of Light-Dark Regulation of the Chloroplast ATP Synthase.

PubMed

Kohzuma, Kaori; Froehlich, John E; Davis, Geoffry A; Temple, Joshua A; Minhas, Deepika; Dhingra, Amit; Cruz, Jeffrey A; Kramer, David M

2017-01-01

The chloroplast ATP synthase catalyzes the light-driven synthesis of ATP and is activated in the light and inactivated in the dark by redox-modulation through the thioredoxin system. It has been proposed that this down-regulation is important for preventing wasteful hydrolysis of ATP in the dark. To test this proposal, we compared the effects of extended dark exposure in Arabidopsis lines expressing the wild-type and mutant forms of ATP synthase that are redox regulated or constitutively active. In contrast to the predictions of the model, we observed that plants with wild-type redox regulation lost photosynthetic capacity rapidly in darkness, whereas those expressing redox-insensitive form were far more stable. To explain these results, we propose that in wild-type plants, down-regulation of ATP synthase inhibits ATP hydrolysis, leading to dissipation of thylakoid proton motive force (pmf) and subsequent inhibition of protein transport across the thylakoid through the twin arginine transporter (Tat)-dependent and Sec-dependent import pathways, resulting in the selective loss of specific protein complexes. By contrast, in mutants with a redox-insensitive ATP synthase, pmf is maintained by ATP hydrolysis, thus allowing protein transport to maintain photosynthetic activities for extended periods in the dark. Hence, a basal level of Tat-dependent, as well as, Sec-dependent import activity, in the dark helps replenishes certain components of the photosynthetic complexes and thereby aids in maintaining overall complex activity. However, the influence of a dark pmf on thylakoid protein import, by itself, could not explain all the effects we observed in this study. For example, we also observed in wild type plants a large transient buildup of thylakoid pmf and nonphotochemical exciton quenching upon sudden illumination of dark adapted plants. Therefore, we conclude that down-regulation of the ATP synthase is probably not related to preventing loss of ATP per se . Instead

The Role of Light–Dark Regulation of the Chloroplast ATP Synthase

PubMed Central

Kohzuma, Kaori; Froehlich, John E.; Davis, Geoffry A.; Temple, Joshua A.; Minhas, Deepika; Dhingra, Amit; Cruz, Jeffrey A.; Kramer, David M.

2017-01-01

The chloroplast ATP synthase catalyzes the light-driven synthesis of ATP and is activated in the light and inactivated in the dark by redox-modulation through the thioredoxin system. It has been proposed that this down-regulation is important for preventing wasteful hydrolysis of ATP in the dark. To test this proposal, we compared the effects of extended dark exposure in Arabidopsis lines expressing the wild-type and mutant forms of ATP synthase that are redox regulated or constitutively active. In contrast to the predictions of the model, we observed that plants with wild-type redox regulation lost photosynthetic capacity rapidly in darkness, whereas those expressing redox-insensitive form were far more stable. To explain these results, we propose that in wild-type plants, down-regulation of ATP synthase inhibits ATP hydrolysis, leading to dissipation of thylakoid proton motive force (pmf) and subsequent inhibition of protein transport across the thylakoid through the twin arginine transporter (Tat)-dependent and Sec-dependent import pathways, resulting in the selective loss of specific protein complexes. By contrast, in mutants with a redox-insensitive ATP synthase, pmf is maintained by ATP hydrolysis, thus allowing protein transport to maintain photosynthetic activities for extended periods in the dark. Hence, a basal level of Tat-dependent, as well as, Sec-dependent import activity, in the dark helps replenishes certain components of the photosynthetic complexes and thereby aids in maintaining overall complex activity. However, the influence of a dark pmf on thylakoid protein import, by itself, could not explain all the effects we observed in this study. For example, we also observed in wild type plants a large transient buildup of thylakoid pmf and nonphotochemical exciton quenching upon sudden illumination of dark adapted plants. Therefore, we conclude that down-regulation of the ATP synthase is probably not related to preventing loss of ATP per se. Instead

The Role of Light–Dark Regulation of the Chloroplast ATP Synthase

DOE PAGES

Kohzuma, Kaori; Froehlich, John E.; Davis, Geoffry A.; ...

2017-07-24

The chloroplast ATP synthase catalyzes the light-driven synthesis of ATP and is activated in the light and inactivated in the dark by redox-modulation through the thioredoxin system. It has been proposed that this down-regulation is important for preventing wasteful hydrolysis of ATP in the dark. To test this proposal, we compared the effects of extended dark exposure in Arabidopsis lines expressing the wild-type and mutant forms of ATP synthase that are redox regulated or constitutively active. In contrast to the predictions of the model, we observed that plants with wild-type redox regulation lost photosynthetic capacity rapidly in darkness, whereas thosemore » expressing redox-insensitive form were far more stable. To explain these results, we propose that in wild-type plants, down-regulation of ATP synthase inhibits ATP hydrolysis, leading to dissipation of thylakoid proton motive force (pmf) and subsequent inhibition of protein transport across the thylakoid through the twin arginine transporter (Tat)-dependent and Secdependent import pathways, resulting in the selective loss of specific protein complexes. By contrast, in mutants with a redox-insensitive ATP synthase, pmf is maintained by ATP hydrolysis, thus allowing protein transport to maintain photosynthetic activities for extended periods in the dark. Hence, a basal level of Tat-dependent, as well as, Sec-dependent import activity, in the dark helps replenishes certain components of the photosynthetic complexes and thereby aids in maintaining overall complex activity. But, the influence of a dark pmf on thylakoid protein import, by itself, could not explain all the effects we observed in this study. For example, we also observed in wild type plants a large transient buildup of thylakoid pmf and nonphotochemical exciton quenching upon sudden illumination of dark adapted plants. Thus, we conclude that down-regulation of the ATP synthase is probably not related to preventing loss of ATP per se. Instead

The Role of Light–Dark Regulation of the Chloroplast ATP Synthase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kohzuma, Kaori; Froehlich, John E.; Davis, Geoffry A.

The chloroplast ATP synthase catalyzes the light-driven synthesis of ATP and is activated in the light and inactivated in the dark by redox-modulation through the thioredoxin system. It has been proposed that this down-regulation is important for preventing wasteful hydrolysis of ATP in the dark. To test this proposal, we compared the effects of extended dark exposure in Arabidopsis lines expressing the wild-type and mutant forms of ATP synthase that are redox regulated or constitutively active. In contrast to the predictions of the model, we observed that plants with wild-type redox regulation lost photosynthetic capacity rapidly in darkness, whereas thosemore » expressing redox-insensitive form were far more stable. To explain these results, we propose that in wild-type plants, down-regulation of ATP synthase inhibits ATP hydrolysis, leading to dissipation of thylakoid proton motive force (pmf) and subsequent inhibition of protein transport across the thylakoid through the twin arginine transporter (Tat)-dependent and Secdependent import pathways, resulting in the selective loss of specific protein complexes. By contrast, in mutants with a redox-insensitive ATP synthase, pmf is maintained by ATP hydrolysis, thus allowing protein transport to maintain photosynthetic activities for extended periods in the dark. Hence, a basal level of Tat-dependent, as well as, Sec-dependent import activity, in the dark helps replenishes certain components of the photosynthetic complexes and thereby aids in maintaining overall complex activity. But, the influence of a dark pmf on thylakoid protein import, by itself, could not explain all the effects we observed in this study. For example, we also observed in wild type plants a large transient buildup of thylakoid pmf and nonphotochemical exciton quenching upon sudden illumination of dark adapted plants. Thus, we conclude that down-regulation of the ATP synthase is probably not related to preventing loss of ATP per se. Instead

ATP during early bladder stretch is important for urgency in detrusor overactivity patients.

PubMed

Cheng, Y; Mansfield, K J; Allen, W; Chess-Williams, R; Burcher, E; Moore, K H

2014-01-01

ATP is an important mediator of urgency in women with detrusor overactivity (DO). In order to understand how different degrees of bladder stretch elicited ATP release in DO patients compared with controls, sequential aliquots were collected during cystometry and ATP release was measured at each degree of bladder filling, in female patients with DO and controls. In both DO and control groups, ATP release was induced during bladder filling, suggesting that stretch stimulated further ATP release. However, the luminal ATP concentrations were already high at early filling stage (200 mL), which was even greater than those at the later filling stages (400 mL and maximum cystometric capacity, MCC), indicating that a substantial ATP release has been induced during early filling (200 mL) in both DO and controls. In DO, ATP release at 200 mL was significantly higher in those with low first desire to void (FDV) (≤ 200 mL) than in those with higher FDV (> 200 mL); this may suggest that ATP release at early stretch may play an important role in urgency (early sensation) in DO. ATP concentrations remained unchanged after voiding, suggesting that voiding did not further induce ATP release into intraluminal fluid.

CFTR Cl- channel and CFTR-associated ATP channel: distinct pores regulated by common gates.

PubMed Central

Sugita, M; Yue, Y; Foskett, J K

1998-01-01

The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel that is regulated by phosphorylation of the R domain and ATP hydrolysis at two nucleotide-binding domains (NBDs). It is controversial whether CFTR conducts ATP or whether CFTR might be closely associated with a separate ATP conductance. To characterize ATP channels associated with CFTR, we analyzed Cl- and ATP single channel-currents in excised inside-out membrane patches from MDCK epithelial cells transiently expressing CFTR. With 100 mM ATP in the pipette and 140 mM Cl- in the bath, ATP channels were associated with CFTR Cl- channels in two-thirds of patches that included CFTR. CFTR Cl- channels and CFTR-associated ATP channels had slope conductances of 7.4 pS and 5.2 pS, respectively, and had distinct reversal potentials and sensitivities to channel blockers. CFTR-associated ATP channels exhibited slow gating kinetics that depended on the presence of protein kinase A and cytoplasmic ATP, similar to CFTR Cl- channels. Gating kinetics of the ATP channels as well as the CFTR Cl- channels were similarly affected by non-hydrolyzable ATP analogues and mutations in the CFTR R domain and NBDs. Our results indicate that phosphorylation- and nucleotide-hydrolysis-dependent gating of CFTR is directly involved in gating of an associated ATP channel. However, the permeation pathways for Cl- and ATP are distinct and the ATP conduction pathway is not obligatorily associated with the expression of CFTR. PMID:9463368

Piezo1 regulates mechanotransductive release of ATP from human RBCs.

PubMed

Cinar, Eyup; Zhou, Sitong; DeCourcey, James; Wang, Yixuan; Waugh, Richard E; Wan, Jiandi

2015-09-22

Piezo proteins (Piezo1 and Piezo2) are recently identified mechanically activated cation channels in eukaryotic cells and associated with physiological responses to touch, pressure, and stretch. In particular, human RBCs express Piezo1 on their membranes, and mutations of Piezo1 have been linked to hereditary xerocytosis. To date, however, physiological functions of Piezo1 on normal RBCs remain poorly understood. Here, we show that Piezo1 regulates mechanotransductive release of ATP from human RBCs by controlling the shear-induced calcium (Ca(2+)) influx. We find that, in human RBCs treated with Piezo1 inhibitors or having mutant Piezo1 channels, the amounts of shear-induced ATP release and Ca(2+) influx decrease significantly. Remarkably, a critical extracellular Ca(2+) concentration is required to trigger significant ATP release, but membrane-associated ATP pools in RBCs also contribute to the release of ATP. Our results show how Piezo1 channels are likely to function in normal RBCs and suggest a previously unidentified mechanotransductive pathway in ATP release. Thus, we anticipate that the study will impact broadly on the research of red cells, cellular mechanosensing, and clinical studies related to red cell disorders and vascular disease.

Determination of effects of ambient conditions on aircraft engine emissions engine testing - GTCP 85 APU, TPE 331 turboprop. Volume 1. Final report 20 Dec 1974--20 Dec 1975

DOE Office of Scientific and Technical Information (OSTI.GOV)

Slogar, G.A.

1976-03-01

Full scale engine tests were conducted on a GTCP85-98CK Auxiliary Power Unit and a TPE331-5-251M Turboprop engine. The purpose of this program was to measure exhaust emission of HC, CO, CO/sub 2/, NO/sub x/, and smoke at controlled (temperature, humidity, and pressure) engine inlet conditions. This data along with other available data will provide the data base for the determination of the effects of ambient conditions on gas turbine engines. (GRA)

Application of luciferase assay for ATP to antimicrobial drug susceptibility

NASA Technical Reports Server (NTRS)

Chappelle, E. W.; Picciolo, G. L.; Vellend, H.; Tuttle, S. A.; Barza, M. J.; Weinstein, L. (Inventor)

1977-01-01

The susceptibility of bacteria, particularly those derived from body fluids, to antimicrobial agents is determined in terms of an ATP index measured by culturing a bacterium in a growth medium. The amount of ATP is assayed in a sample of the cultured bacterium by measuring the amount of luminescent light emitted when the bacterial ATP is reacted with a luciferase-luciferin mixture. The sample of the cultured bacterium is subjected to an antibiotic agent. The amount of bacterial adenosine triphosphate is assayed after treatment with the antibiotic by measuring the luminescent light resulting from the reaction. The ATP index is determined from the values obtained from the assay procedures.

Agonist trapped in ATP-binding sites of the P2X2 receptor

PubMed Central

Jiang, Ruotian; Lemoine, Damien; Martz, Adeline; Taly, Antoine; Gonin, Sophie; Prado de Carvalho, Lia; Specht, Alexandre; Grutter, Thomas

2011-01-01

ATP-gated P2X receptors are trimeric ion channels, as recently confirmed by X-ray crystallography. However, the structure was solved without ATP and even though extracellular intersubunit cavities surrounded by conserved amino acid residues previously shown to be important for ATP function were proposed to house ATP, the localization of the ATP sites remains elusive. Here we localize the ATP-binding sites by creating, through a proximity-dependent “tethering” reaction, covalent bonds between a synthesized ATP-derived thiol-reactive P2X2 agonist (NCS-ATP) and single cysteine mutants engineered in the putative binding cavities of the P2X2 receptor. By combining whole-cell and single-channel recordings, we report that NCS-ATP covalently and specifically labels two previously unidentified positions N140 and L186 from two adjacent subunits separated by about 18 Å in a P2X2 closed state homology model, suggesting the existence of at least two binding modes. Tethering reaction at both positions primes subsequent agonist binding, yet with distinct functional consequences. Labeling of one position impedes subsequent ATP function, which results in inefficient gating, whereas tethering of the other position, although failing to produce gating by itself, enhances subsequent ATP function. Our results thus define a large and dynamic intersubunit ATP-binding pocket and suggest that receptors trapped in covalently agonist-bound states differ in their ability to gate the ion channel. PMID:21576497

Agonist trapped in ATP-binding sites of the P2X2 receptor.

PubMed

Jiang, Ruotian; Lemoine, Damien; Martz, Adeline; Taly, Antoine; Gonin, Sophie; Prado de Carvalho, Lia; Specht, Alexandre; Grutter, Thomas

2011-05-31

ATP-gated P2X receptors are trimeric ion channels, as recently confirmed by X-ray crystallography. However, the structure was solved without ATP and even though extracellular intersubunit cavities surrounded by conserved amino acid residues previously shown to be important for ATP function were proposed to house ATP, the localization of the ATP sites remains elusive. Here we localize the ATP-binding sites by creating, through a proximity-dependent "tethering" reaction, covalent bonds between a synthesized ATP-derived thiol-reactive P2X2 agonist (NCS-ATP) and single cysteine mutants engineered in the putative binding cavities of the P2X2 receptor. By combining whole-cell and single-channel recordings, we report that NCS-ATP covalently and specifically labels two previously unidentified positions N140 and L186 from two adjacent subunits separated by about 18 Å in a P2X2 closed state homology model, suggesting the existence of at least two binding modes. Tethering reaction at both positions primes subsequent agonist binding, yet with distinct functional consequences. Labeling of one position impedes subsequent ATP function, which results in inefficient gating, whereas tethering of the other position, although failing to produce gating by itself, enhances subsequent ATP function. Our results thus define a large and dynamic intersubunit ATP-binding pocket and suggest that receptors trapped in covalently agonist-bound states differ in their ability to gate the ion channel.

Dental, Dental Hygiene, and Advanced Dental Students' Use, Knowledge, and Beliefs Regarding Tobacco Products.

PubMed

Shearston, Jenni A; Shah, Krina; Cheng, Eric; Moosvi, Rizvan; Park, Su Hyun; Patel, Naiya; Spielman, Andrew I; Weitzman, Michael L

2017-11-01

Using cigarettes and alternative tobacco products (ATPs) is associated with negative oral health outcomes, and dental health professionals are poised to help patients quit. The aim of this study was to determine dental, dental hygiene, and advanced dental students' use, knowledge, and beliefs about cigarettes and ATPs, including perceptions about their education in tobacco dependence treatment and counseling experience. All 1,783 students enrolled in the dental, dental hygiene, and postdoctoral dental programs at the New York University College of Dentistry were invited to participate in the survey in 2016. A total of 708 students at least partially completed the survey, for a response rate of 39.7%. In the results, 146 of the students (20.1%) reported ever using cigarettes, while 253 (35.7%) reported ever using any ATP. Regarding tobacco use intervention, the students reported they had not received enough training on ATPs, were neutral about cigarettes, and were somewhat confident and not so confident counseling a cigarette smoker or ATP user, respectively. By their fourth year, 77.8% of the dental students reported they had counseled someone to stop smoking cigarettes, but only 40.7% had counseled someone to stop using ATPs. Overall, all groups of students reported feeling more confident and had received more education on interventions for cigarettes than for ATPs (p<0.001). These students reported low confidence in helping people quit tobacco and did not perceive they had received enough training on intervening with patients on use of cigarettes and ATPs. These findings call for a revised tobacco education curriculum for dental, dental hygiene, and advanced dental students, focused on building knowledge and confidence for promoting tobacco dependence treatment.

Bimodal Action of Protons on ATP Currents of Rat PC12 Cells

PubMed Central

Skorinkin, Andrei; Nistri, Andrea; Giniatullin, Rashid

2003-01-01

The mode of action of extracellular protons on ATP-gated P2X2 receptors remains controversial as either enhancement or depression of ATP-mediated currents has been reported. By investigating, at different pH, the electrophysiological effect of ATP on P2X2 receptors and complementing it with receptor modelling, the present study suggests a unified mechanism for both potentiation and inactivation of ATP receptors by protons. Our experiments on patch-clamped PC12 cells showed that, on the same cell, mild acidification potentiated currents induced by low ATP concentrations (<0.1 mM) and attenuated responses to high ATP concentrations (>1 mM) with emergence of current fading and rebound. To clarify the nature of the ATP/H+ interaction, we used the Ding and Sachs's “loop” receptor model which best describes the behavior of such receptors with two open states linked via one inactivated state. No effects by protons could be ascribed to H+-mediated open channel block. However, by assuming that protons facilitated binding of ATP to resting as well as open receptors, the model could closely replicate H+-induced potentiation of currents evoked by low ATP doses plus fading and rebound induced by high ATP doses. The latter phenomenon was due to receptor transition to the inactive state. The present data suggest that the high concentration of protons released with ATP (and catecholamines) from secretory vesicles may allow a dual action of H+ on P2X2 receptors. This condition might also occur on P2X2 receptors of central neurons exposed to low pH during ischemia. PMID:12810852

ATP can be dispensable for prespliceosome formation in yeast

PubMed Central

Perriman, Rhonda; Ares, Manuel

2000-01-01

The first ATP-dependent step in pre-mRNA splicing involves the stable binding of U2 snRNP to form the prespliceosome. We show that a prespliceosome-like complex forms in the absence of ATP in yeast extracts lacking the U2 suppressor protein CUS2. These complexes display the same pre-mRNA and U snRNA requirements as authentic prespliceosomes and can be chased through the splicing pathway, indicating that they are a functional intermediate in the spliceosome assembly pathway. ATP-independent prespliceosome-like complexes are also observed in extracts containing a mutant U2 snRNA. Loss of CUS2 does not bypass the role of PRP5, an RNA helicase family member required for ATP-dependent prespliceosome formation. Genetic interactions between CUS2 and a heat-sensitive prp5 allele parallel those observed between CUS2 and U2, and suggest that CUS2 mediates functional interactions between U2 RNA and PRP5. We propose that CUS2 enforces ATP dependence during formation of the prespliceosome by brokering an interaction between PRP5 and the U2 snRNP that depends on correct U2 RNA structure. PMID:10640279

Large-Scale Advanced Prop-Fan (LAP) pitch change actuator and control design report

NASA Technical Reports Server (NTRS)

Schwartz, R. A.; Carvalho, P.; Cutler, M. J.

1986-01-01

In recent years, considerable attention has been directed toward improving aircraft fuel consumption. Studies have shown that the high inherent efficiency previously demonstrated by low speed turboprop propulsion systems may now be extended to today's higher speed aircraft if advanced high-speed propeller blades having thin airfoils and aerodynamic sweep are utilized. Hamilton Standard has designed a 9-foot diameter single-rotation Large-Scale Advanced Prop-Fan (LAP) which will be tested on a static test stand, in a high speed wind tunnel and on a research aircraft. The major objective of this testing is to establish the structural integrity of large-scale Prop-Fans of advanced construction in addition to the evaluation of aerodynamic performance and aeroacoustic design. This report describes the operation, design features and actual hardware of the (LAP) Prop-Fan pitch control system. The pitch control system which controls blade angle and propeller speed consists of two separate assemblies. The first is the control unit which provides the hydraulic supply, speed governing and feather function for the system. The second unit is the hydro-mechanical pitch change actuator which directly changes blade angle (pitch) as scheduled by the control.

NMR-invisible ATP in heart: fact or fiction?

PubMed

Bak, M I; Ingwall, J S

1992-06-01

31P-nuclear magnetic resonance (31P-NMR) spectroscopy is widely used to monitor sequential changes in the nucleoside triphosphate (NTP) pool in intact tissues. Recently, the validity of this technique to quantitate incremental changes in ATP in heart has been challenged. Accordingly, we compared NTP measured by 31P-NMR and by chemical techniques in isolated isovolumic rat hearts at 16 and 56 min of oxygenated perfusion and in hearts subjected to 28 min of hypoxia, with or without 28 min of reoxygenation, and 12 or 28 min of ischemia, with or without 28 min of reperfusion. NTP content was calculated from 31P-NMR spectra using an external standard. At the end of each protocol the heart was freeze-clamped, and NTP and ATP contents were determined by chemical assay. After 16 min of normoxic perfusion the values for NTP and ATP contents measured by both methods in the same hearts were indistinguishable. Results from all seven experimental conditions show no significant difference between methods (P = 0.262). Thus both methods detect the same incremental change in NTP and ATP.

ATP-induced noncooperative thermal unfolding of hen lysozyme

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Honglin; Yin, Peidong; He, Shengnan

To understand the role of ATP underlying the enhanced amyloidosis of hen egg white lysozyme (HEWL), the synchrotron radiation circular dichroism, combined with tryptophan fluorescence, dynamic light-scattering, and differential scanning calorimetry, is used to examine the alterations of the conformation and thermal unfolding pathway of the HEWL in the presence of ATP, Mg{sup 2+}-ATP, ADP, AMP, etc. It is revealed that the binding of ATP to HEWL through strong electrostatic interaction changes the secondary structures of HEWL and makes the exposed residue W62 move into hydrophobic environments. This alteration of W62 decreases the {beta}-domain stability of HEWL, induces a noncooperativemore » unfolding of the secondary structures, and produces a partially unfolded intermediate. This intermediate containing relatively rich {alpha}-helix and less {beta}-sheet structures has a great tendency to aggregate. The results imply that the ease of aggregating of HEWL is related to the extent of denaturation of the amyloidogenic region, rather than the electrostatic neutralizing effect or monomeric {beta}-sheet enriched intermediate.« less

Failure of the Cystic Fibrosis Transmembrane Conductance Regulator to Conduct ATP

NASA Astrophysics Data System (ADS)

Reddy, M. M.; Quinton, P. M.; Haws, C.; Wine, J. J.; Grygorczyk, R.; Tabcharani, J. A.; Hanrahan, J. W.; Gunderson, K. L.; Kopito, R. R.

1996-03-01

The cystic fibrosis transmembrane conductance regulator (CFTR) is chloride ion channel regulated by protein kinase A and adenosine triphosphate (ATP). Loss of CFTR-mediated chloride ion conductance from the apical plasma membrane of epithelial cells is a primary physiological lesion in cystic fibrosis. CFTR has also been suggested to function as an ATP channel, although the size of the ATP anion is much larger than the estimated size of the CFTR pore. ATP was not conducted through CFTR in intact organs, polarized human lung cell lines, stably transfected mammalian cell lines, or planar lipid bilayers reconstituted with CFTR protein. These findings suggest that ATP permeation through the CFTR is unlikely to contribute to the normal function of CFTR or to the pathogenesis of cystic fibrosis.

Inflammation Promotes Airway Epithelial ATP Release via Calcium-Dependent Vesicular Pathways

PubMed Central

Okada, Seiko F.; Ribeiro, Carla M. P.; Sesma, Juliana I.; Seminario-Vidal, Lucia; Abdullah, Lubna H.; van Heusden, Catharina; Lazarowski, Eduardo R.

2013-01-01

ATP in airway surface liquid (ASL) controls mucociliary clearance functions via the activation of airway epithelial purinergic receptors. However, abnormally elevated ATP levels have been reported in inflamed airways, suggesting that excessive ATP in ASL contributes to airway inflammation. Despite these observations, little is known about the mechanisms of ATP accumulation in the ASL covering inflamed airways. In this study, links between cystic fibrosis (CF)–associated airway inflammation and airway epithelial ATP release were investigated. Primary human bronchial epithelial (HBE) cells isolated from CF lungs exhibited enhanced IL-8 secretion after 6 to 11 days, but not 28 to 35 days, in culture, compared with normal HBE cells. Hypotonic cell swelling–promoted ATP release was increased in 6- to 11-day-old CF HBE cells compared with non-CF HBE cells, but returned to normal values after 28 to 35 days in culture. The exposure of non-CF HBE cells to airway secretions isolated from CF lungs, namely, sterile supernatants of mucopurulent material (SMM), also caused enhanced IL-8 secretion and increased ATP release. The SMM-induced increase in ATP release was sensitive to Ca2+ chelation and vesicle trafficking/exocytosis inhibitors, but not to pannexin inhibition. Transcript levels of the vesicular nucleotide transporter, but not pannexin 1, were up-regulated after SMM exposure. SMM-treated cultures displayed increased basal mucin secretion, but mucin secretion was not enhanced in response to hypotonic challenge after the exposure of cells to either vehicle or SMM. We propose that CF airway inflammation up-regulates the capacity of airway epithelia to release ATP via Ca2+-dependent vesicular mechanisms not associated with mucin granule secretion. PMID:23763446

Probing the ATP site of GRP78 with nucleotide triphosphate analogs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hughes, Scott J.; Antoshchenko, Tetyana; Chen, Yun

GRP78, a member of the ER stress protein family, can relocate to the surface of cancer cells, playing key roles in promoting cell proliferation and metastasis. GRP78 consists of two major functional domains: the ATPase and protein/peptide-binding domains. The protein/peptide-binding domain of cell-surface GRP78 has served as a novel functional receptor for delivering cytotoxic agents (e.g., a apoptosis-inducing peptide or taxol) across the cell membrane. Here, we report our study on the ATPase domain of GRP78 (GRP78 ATPase), whose potential as a transmembrane delivery system of cytotoxic agents (e.g., ATP-based nucleotide triphosphate analogs) remains unexploited. As the binding of ligandsmore » (ATP analogs) to a receptor (GRP78 ATPase) is a pre-requisite for internalization, we determined the binding affinities and modes of GRP78 ATPase for ADP, ATP and several ATP analogs using surface plasmon resonance and x-ray crystallography. The tested ATP analogs contain one of the following modifications: the nitrogen at the adenine ring 7-position to a carbon atom (7-deazaATP), the oxygen at the beta-gamma bridge position to a carbon atom (AMPPCP), or the removal of the 2'-OH group (2'-deoxyATP). We found that 7-deazaATP displays an affinity and a binding mode that resemble those of ATP regardless of magnesium ion (Mg ++) concentration, suggesting that GRP78 is tolerant to modifications at the 7-position. By comparison, AMPPCP's binding affinity was lower than ATP and Mg ++-dependent, as the removal of Mg ++ nearly abolished binding to GRP78 ATPase. The AMPPCP-Mg ++ structure showed evidence for the critical role of Mg ++ in AMPPCP binding affinity, suggesting that while GRP78 is sensitive to modifications at the β-γ bridge position, these can be tolerated in the presence of Mg ++. Furthermore, 2'-deoxyATP's binding affinity was significantly lower than those for all other nucleotides tested, even in the presence of Mg ++. The 2'-deoxyATP structure showed the

Probing the ATP site of GRP78 with nucleotide triphosphate analogs

DOE PAGES

Hughes, Scott J.; Antoshchenko, Tetyana; Chen, Yun; ...

2016-05-04

GRP78, a member of the ER stress protein family, can relocate to the surface of cancer cells, playing key roles in promoting cell proliferation and metastasis. GRP78 consists of two major functional domains: the ATPase and protein/peptide-binding domains. The protein/peptide-binding domain of cell-surface GRP78 has served as a novel functional receptor for delivering cytotoxic agents (e.g., a apoptosis-inducing peptide or taxol) across the cell membrane. Here, we report our study on the ATPase domain of GRP78 (GRP78 ATPase), whose potential as a transmembrane delivery system of cytotoxic agents (e.g., ATP-based nucleotide triphosphate analogs) remains unexploited. As the binding of ligandsmore » (ATP analogs) to a receptor (GRP78 ATPase) is a pre-requisite for internalization, we determined the binding affinities and modes of GRP78 ATPase for ADP, ATP and several ATP analogs using surface plasmon resonance and x-ray crystallography. The tested ATP analogs contain one of the following modifications: the nitrogen at the adenine ring 7-position to a carbon atom (7-deazaATP), the oxygen at the beta-gamma bridge position to a carbon atom (AMPPCP), or the removal of the 2'-OH group (2'-deoxyATP). We found that 7-deazaATP displays an affinity and a binding mode that resemble those of ATP regardless of magnesium ion (Mg ++) concentration, suggesting that GRP78 is tolerant to modifications at the 7-position. By comparison, AMPPCP's binding affinity was lower than ATP and Mg ++-dependent, as the removal of Mg ++ nearly abolished binding to GRP78 ATPase. The AMPPCP-Mg ++ structure showed evidence for the critical role of Mg ++ in AMPPCP binding affinity, suggesting that while GRP78 is sensitive to modifications at the β-γ bridge position, these can be tolerated in the presence of Mg ++. Furthermore, 2'-deoxyATP's binding affinity was significantly lower than those for all other nucleotides tested, even in the presence of Mg ++. The 2'-deoxyATP structure showed the

ATP synthase promotes germ cell differentiation independent of oxidative phosphorylation

PubMed Central

Teixeira, Felipe K.; Sanchez, Carlos G.; Hurd, Thomas R.; Seifert, Jessica R. K.; Czech, Benjamin; Preall, Jonathan B.; Hannon, Gregory J.; Lehmann, Ruth

2015-01-01

The differentiation of stem cells is a tightly regulated process essential for animal development and tissue homeostasis. Through this process, attainment of new identity and function is achieved by marked changes in cellular properties. Intrinsic cellular mechanisms governing stem cell differentiation remain largely unknown, in part because systematic forward genetic approaches to the problem have not been widely used1,2. Analysing genes required for germline stem cell differentiation in the Drosophila ovary, we find that the mitochondrial ATP synthase plays a critical role in this process. Unexpectedly, the ATP synthesizing function of this complex was not necessary for differentiation, as knockdown of other members of the oxidative phosphorylation system did not disrupt the process. Instead, the ATP synthase acted to promote the maturation of mitochondrial cristae during differentiation through dimerization and specific upregulation of the ATP synthase complex. Taken together, our results suggest that ATP synthase-dependent crista maturation is a key developmental process required for differentiation independent of oxidative phosphorylation. PMID:25915123

Synergic effects of mycoplasmal lipopeptides and extracellular ATP on activation of macrophages.

PubMed

Into, Takeshi; Fujita, Mari; Okusawa, Tsugumi; Hasebe, Akira; Morita, Manabu; Shibata, Ken-Ichiro

2002-07-01

Mycoplasmal lipopeptides S-(2,3-bispalmitoyloxypropyl)-CGDPKHSPKSF and S-(2,3-bispalmitoyloxypropyl)-CGNNDESNISFKEK activated a monocytic cell line, THP-1 cells, to produce tumor necrosis factor alpha. The activity of the lipopeptides was augmented by ATP in a dose-dependent manner. In addition, the level of expression of mRNAs for tumor necrosis factor alpha and interleukin-1 beta, -6, and -8 was also upregulated by the lipopeptides and/or extracellular ATP, but that of interleukin-10 was not. The P2X purinergic receptor antagonists pyridoxal phosphate 6-azophenyl 2',4'-disulfonic acid and periodate-oxidized ATP suppressed the activity of ATP to augment the activation of THP-1 cells by the lipopeptides, suggesting that P2X receptors play important roles in the activity of ATP. The nuclear factor kappa B inhibitor dexamethasone also suppressed the activity, suggesting that the activity of ATP is dependent upon the nuclear factor kappa B. Thus, these results suggest that the interaction of extracellular ATP with the P2X receptors is attributed to the activity of ATP to augment the activation of THP-1 cells by mycoplasmal lipopeptides.

Synergic Effects of Mycoplasmal Lipopeptides and Extracellular ATP on Activation of Macrophages

PubMed Central

Into, Takeshi; Fujita, Mari; Okusawa, Tsugumi; Hasebe, Akira; Morita, Manabu; Shibata, Ken-Ichiro

2002-01-01

Mycoplasmal lipopeptides S-(2,3-bispalmitoyloxypropyl)-CGDPKHSPKSF and S-(2,3-bispalmitoyloxypropyl)-CGNNDESNISFKEK activated a monocytic cell line, THP-1 cells, to produce tumor necrosis factor alpha. The activity of the lipopeptides was augmented by ATP in a dose-dependent manner. In addition, the level of expression of mRNAs for tumor necrosis factor alpha and interleukin-1β, -6, and -8 was also upregulated by the lipopeptides and/or extracellular ATP, but that of interleukin-10 was not. The P2X purinergic receptor antagonists pyridoxal phosphate 6-azophenyl 2′,4′-disulfonic acid and periodate-oxidized ATP suppressed the activity of ATP to augment the activation of THP-1 cells by the lipopeptides, suggesting that P2X receptors play important roles in the activity of ATP. The nuclear factor κB inhibitor dexamethasone also suppressed the activity, suggesting that the activity of ATP is dependent upon the nuclear factor κB. Thus, these results suggest that the interaction of extracellular ATP with the P2X receptors is attributed to the activity of ATP to augment the activation of THP-1 cells by mycoplasmal lipopeptides. PMID:12065499

Laser beam propagation through a full scale aircraft turboprop engine exhaust

NASA Astrophysics Data System (ADS)

Henriksson, Markus; Gustafsson, Ove; Sjöqvist, Lars; Seiffer, Dirk; Wendelstein, Norbert

2010-10-01

The exhaust from engines introduces zones of extreme turbulence levels in local environments around aircraft. This may disturb the performance of aircraft mounted optical and laser systems. The turbulence distortion will be especially devastating for optical missile warning and laser based DIRCM systems used to protect manoeuvring aircraft against missile attacks, situations where the optical propagation path may come close to the engine exhaust. To study the extent of the turbulence zones caused by the engine exhaust and the strength of the effects on optical propagation through these zones a joint trial between Germany, the Netherlands, Sweden and the United Kingdom was performed using a medium sized military turboprop transport aircraft tethered to the ground at an airfield. This follows on earlier trials performed on a down-scaled jet-engine test rig. Laser beams were propagated along the axis of the aircraft at different distances relative to the engine exhaust and the spatial beam profiles and intensity scintillations were recorded with cameras and photodiodes. A second laser beam path was directed from underneath the loading ramp diagonally past one of the engines. The laser wavelengths used were 1.5 and 3.6 μm. In addition to spatial beam profile distortions temporal effects were investigated. Measurements were performed at different propeller speeds and at different distances from exhaust nozzle to the laser path. Significant increases in laser beam wander and long term beam radius were observed with the engine running. Corresponding increases were also registered in the scintillation index and the temporal fluctuations of the instantaneous power collected by the detector.

ATP as a biomarker of viable microorganisms in clean-room facilities

NASA Technical Reports Server (NTRS)

Venkateswaran, Kasthuri; Hattori, Noriaki; La Duc, Myron T.; Kern, Roger

2003-01-01

A new firefly luciferase bioluminescence assay method that differentiates free extracellular ATP (dead cells, etc.) from intracellular ATP (viable microbes) was used to determine the viable microbial cleanliness of various clean-room facilities. For comparison, samples were taken from both clean-rooms, where the air was filtered to remove particles >0.5 microm, and ordinary rooms with unfiltered air. The intracellular ATP was determined after enzymatically degrading the sample's free ATP. Also for comparison, cultivable microbial populations were counted on nutrient-rich trypticase soy agar (TSA) plates. Both the cultivable and ATP-based determinations indicate that the microbial burden was lower in clean-room facilities than in ordinary rooms. However, there was no direct correlation between the two sets of measurements because the two assays measured very different populations. A large fraction of the samples yielded no colony formers on TSA, but were positive for intracellular ATP. Subsequently, genomic DNA was isolated directly from selected samples and 16S rDNA fragments were cloned and sequenced, identifying nearest neighbors, many of which are known to be noncultivable in the media employed. It was concluded that viable microbial contamination can be reliably monitored by measurement of intracellular ATP, and that this method may be considered superior to cultivable colony counts due to its speed and its ability to report the presence of viable but noncultivable organisms. When the detection of nonviable microbes is of interest, the ATP assay can be supplemented with DNA analysis.

ATP as a biomarker of viable microorganisms in clean-room facilities.

PubMed

Venkateswaran, Kasthuri; Hattori, Noriaki; La Duc, Myron T; Kern, Roger

2003-03-01

A new firefly luciferase bioluminescence assay method that differentiates free extracellular ATP (dead cells, etc.) from intracellular ATP (viable microbes) was used to determine the viable microbial cleanliness of various clean-room facilities. For comparison, samples were taken from both clean-rooms, where the air was filtered to remove particles >0.5 microm, and ordinary rooms with unfiltered air. The intracellular ATP was determined after enzymatically degrading the sample's free ATP. Also for comparison, cultivable microbial populations were counted on nutrient-rich trypticase soy agar (TSA) plates. Both the cultivable and ATP-based determinations indicate that the microbial burden was lower in clean-room facilities than in ordinary rooms. However, there was no direct correlation between the two sets of measurements because the two assays measured very different populations. A large fraction of the samples yielded no colony formers on TSA, but were positive for intracellular ATP. Subsequently, genomic DNA was isolated directly from selected samples and 16S rDNA fragments were cloned and sequenced, identifying nearest neighbors, many of which are known to be noncultivable in the media employed. It was concluded that viable microbial contamination can be reliably monitored by measurement of intracellular ATP, and that this method may be considered superior to cultivable colony counts due to its speed and its ability to report the presence of viable but noncultivable organisms. When the detection of nonviable microbes is of interest, the ATP assay can be supplemented with DNA analysis.

Large-Scale Advanced Prop-Fan (LAP)

NASA Technical Reports Server (NTRS)

Degeorge, C. L.

1988-01-01

In recent years, considerable attention has been directed toward improving aircraft fuel efficiency. Analytical studies and research with wind tunnel models have demonstrated that the high inherent efficiency of low speed turboprop propulsion systems may now be extended to the Mach .8 flight regime of today's commercial airliners. This can be accomplished with a propeller, employing a large number of thin highly swept blades. The term Prop-Fan has been coined to describe such a propulsion system. In 1983 the NASA-Lewis Research Center contracted with Hamilton Standard to design, build and test a near full scale Prop-Fan, designated the Large Scale Advanced Prop-Fan (LAP). This report provides a detailed description of the LAP program. The assumptions and analytical procedures used in the design of Prop-Fan system components are discussed in detail. The manufacturing techniques used in the fabrication of the Prop-Fan are presented. Each of the tests run during the course of the program are also discussed and the major conclusions derived from them stated.

An autocrine ATP release mechanism regulates basal ciliary activity in airway epithelium.

PubMed

Droguett, Karla; Rios, Mariana; Carreño, Daniela V; Navarrete, Camilo; Fuentes, Christian; Villalón, Manuel; Barrera, Nelson P

2017-07-15

Extracellular ATP, in association with [Ca 2+ ] i regulation, is required to maintain basal ciliary beat frequency. Increasing extracellular ATP levels increases ciliary beating in airway epithelial cells, maintaining a sustained response by inducing the release of additional ATP. Extracellular ATP levels in the millimolar range, previously associated with pathophysiological conditions of the airway epithelium, produce a transient arrest of ciliary activity. The regulation of ciliary beat frequency is dependent on ATP release by hemichannels (connexin/pannexin) and P2X receptor activation, the blockage of which may even stop ciliary movement. The force exerted by cilia, measured by atomic force microscopy, is reduced following extracellular ATP hydrolysis. This result complements the current understanding of the ciliary beating regulatory mechanism, with special relevance to inflammatory diseases of the airway epithelium that affect mucociliary clearance. Extracellular nucleotides, including ATP, are locally released by the airway epithelium and stimulate ciliary activity in a [Ca 2+ ] i -dependent manner after mechanical stimulation of ciliated cells. However, it is unclear whether the ATP released is involved in regulating basal ciliary activity and mediating changes in ciliary activity in response to chemical stimulation. In the present study, we evaluated ciliary beat frequency (CBF) and ciliary beating forces in primary cultures from mouse tracheal epithelium, using videomicroscopy and atomic force microscopy (AFM), respectively. Extracellular ATP levels and [Ca 2+ ] i were measured by luminometric and fluorimetric assays, respectively. Uptake of ethidium bromide was measured to evaluate hemichannel functionality. We show that hydrolysis of constitutive extracellular ATP levels with apyrase (50 U ml -1 ) reduced basal CBF by 45% and ciliary force by 67%. The apyrase effect on CBF was potentiated by carbenoxolone, a hemichannel inhibitor, and oxidized ATP, an

ATP-Binding Cassette Proteins: Towards a Computational View of Mechanism

NASA Astrophysics Data System (ADS)

Liao, Jielou

2004-03-01

Many large machine proteins can generate mechanical force and undergo large-scale conformational changes (LSCC) to perform varying biological tasks in living cells by utilizing ATP. Important examples include ATP-binding cassette (ABC) transporters. They are membrane proteins that couple ATP binding and hydrolysis to the translocation of substrates across membranes [1]. To interpret how the mechanical force generated by ATP binding and hydrolysis is propagated, a coarse-grained ATP-dependent harmonic network model (HNM) [2,3] is applied to the ABC protein, BtuCD. This protein machine transports vitamin B12 across membranes. The analysis shows that subunits of the protein move against each other in a concerted manner. The lowest-frequency modes of the BtuCD protein are found to link the functionally critical domains, and are suggested to be responsible for large-scale ATP-coupled conformational changes. [1] K. P. Locher, A. T. Lee and D. C. Rees. Science 296, 1091-1098 (2002). [2] Atilgan, A. R., S. R. Durell, R. L. Jernigan, M. C. Demirel, O. Keskin, and I. Bahar. Biophys. J. 80, 505-515(2002); M. M Tirion, Phys. Rev. Lett. 77, 1905-1908 (1996). [3] J. -L. Liao and D. N. Beratan, 2003, to be published.

Processing mechanics of alternate twist ply (ATP) yarn technology

NASA Astrophysics Data System (ADS)

Elkhamy, Donia Said

Ply yarns are important in many textile manufacturing processes and various applications. The primary process used for producing ply yarns is cabling. The speed of cabling is limited to about 35m/min. With the world's increasing demands of ply yarn supply, cabling is incompatible with today's demand activated manufacturing strategies. The Alternate Twist Ply (ATP) yarn technology is a relatively new process for producing ply yarns with improved productivity and flexibility. This technology involves self plying of twisted singles yarn to produce ply yarn. The ATP process can run more than ten times faster than cabling. To implement the ATP process to produce ply yarns there are major quality issues; uniform Twist Profile and yarn Twist Efficiency. The goal of this thesis is to improve these issues through process modeling based on understanding the physics and processing mechanics of the ATP yarn system. In our study we determine the main parameters that control the yarn twist profile. Process modeling of the yarn twist across different process zones was done. A computational model was designed to predict the process parameters required to achieve a square wave twist profile. Twist efficiency, a measure of yarn torsional stability and bulk, is determined by the ratio of ply yarn twist to singles yarn twist. Response Surface Methodology was used to develop the processing window that can reproduce ATP yarns with high twist efficiency. Equilibrium conditions of tensions and torques acting on the yarns at the self ply point were analyzed and determined the pathway for achieving higher twist efficiency. Mechanistic modeling relating equilibrium conditions to the twist efficiency was developed. A static tester was designed to zoom into the self ply zone of the ATP yarn. A computer controlled, prototypic ATP machine was constructed and confirmed the mechanistic model results. Optimum parameters achieving maximum twist efficiency were determined in this study. The

ATP monitoring technology for microbial growth control in potable water systems

NASA Astrophysics Data System (ADS)

Whalen, Patrick A.; Whalen, Philip J.; Cairns, James E.

2006-05-01

ATP (Adenosine Triphosphate) is the primary energy transfer molecule present in all living biological cells on Earth. ATP cannot be produced or maintained by anything but a living organism, and as such, its measurement is a direct indication of biological activity. The main advantage of ATP as a biological indicator is the speed of the analysis - from collecting the sample to obtaining the result, only minutes are required. The technology to measure ATP is already widely utilized to verify disinfection efficacy in the food industry and is also commonly applied in industrial water processes such as cooling water systems to monitor microbial growth and biocide applications. Research has indicated that ATP measurement technology can also play a key role in such important industries as potable water distribution and biological wastewater treatment. As will be detailed in this paper, LuminUltra Technologies has developed and applied ATP measurement technologies designed for any water type, and as such can provide a method to rapidly and accurately determine the level of biological activity in drinking water supplies. Because of its speed and specificity to biological activity, ATP measurement can play a key role in defending against failing drinking water quality, including those encountered during routine operation and also bioterrorism.

Recent advances in enzyme extraction strategies: A comprehensive review.

PubMed

Nadar, Shamraja S; Pawar, Rohini G; Rathod, Virendra K

2017-08-01

The increasing interest of industrial enzymes demands for development of new downstream strategies for maximizing enzyme recovery. The significant efforts have been focused on the development of newly adapted technologies to purify enzymes in catalytically active form. Recently, an aqueous two phase system (ATPS) is emerged as powerful tools for efficient extraction and purification of enzymes due to their versatility, lower cost, process integration capability and easy scale-up. The present review gives an overview of effect of parameters such as tie line length, pH, neutral salts, properties of polymer and salt involved in traditional polymer/polymer and polymer/salt ATPS for enzyme recovery. Further, advanced ATPS have been developed based on alcohols, surfactants, micellar compounds to avoid tedious recovery steps for getting desired enzyme. In order to improve the selectivity and efficiency of ATPS, recent approaches of conventional ATPS combined with different techniques like affinity ligands, ionic liquids, thermoseparating polymers and microfluidic device based ATPS have been reviewed. Moreover, three phase partitioning is also highlighted for enzymes enrichment as a blooming technology for efficiently integrated bioseparation techniques. At the end, it includes an overview of CLEAs technology and organic-inorganic nanoflowers preparation as novel strategies for simultaneous extraction, purification and immobilization of enzymes. Copyright © 2017 Elsevier B.V. All rights reserved.

31P magnetization transfer measurements of Pi→ATP flux in exercising human muscle

PubMed Central

Savage, David B.; Williams, Guy B.; Porter, David; Carpenter, T. Adrian; Brindle, Kevin M.; Kemp, Graham J.

2016-01-01

Fundamental criticisms have been made over the use of 31P magnetic resonance spectroscopy (MRS) magnetization transfer estimates of inorganic phosphate (Pi)→ATP flux (VPi-ATP) in human resting skeletal muscle for assessing mitochondrial function. Although the discrepancy in the magnitude of VPi-ATP is now acknowledged, little is known about its metabolic determinants. Here we use a novel protocol to measure VPi-ATP in human exercising muscle for the first time. Steady-state VPi-ATP was measured at rest and over a range of exercise intensities and compared with suprabasal oxidative ATP synthesis rates estimated from the initial rates of postexercise phosphocreatine resynthesis (VATP). We define a surplus Pi→ATP flux as the difference between VPi-ATP and VATP. The coupled reactions catalyzed by the glycolytic enzymes GAPDH and phosphoglycerate kinase (PGK) have been shown to catalyze measurable exchange between ATP and Pi in some systems and have been suggested to be responsible for this surplus flux. Surplus VPi-ATP did not change between rest and exercise, even though the concentrations of Pi and ADP, which are substrates for GAPDH and PGK, respectively, increased as expected. However, involvement of these enzymes is suggested by correlations between absolute and surplus Pi→ATP flux, both at rest and during exercise, and the intensity of the phosphomonoester peak in the 31P NMR spectrum. This peak includes contributions from sugar phosphates in the glycolytic pathway, and changes in its intensity may indicate changes in downstream glycolytic intermediates, including 3-phosphoglycerate, which has been shown to influence the exchange between ATP and Pi catalyzed by GAPDH and PGK. PMID:26744504

31P magnetization transfer measurements of Pi→ATP flux in exercising human muscle.

PubMed

Sleigh, Alison; Savage, David B; Williams, Guy B; Porter, David; Carpenter, T Adrian; Brindle, Kevin M; Kemp, Graham J

2016-03-15

Fundamental criticisms have been made over the use of (31)P magnetic resonance spectroscopy (MRS) magnetization transfer estimates of inorganic phosphate (Pi)→ATP flux (VPi-ATP) in human resting skeletal muscle for assessing mitochondrial function. Although the discrepancy in the magnitude of VPi-ATP is now acknowledged, little is known about its metabolic determinants. Here we use a novel protocol to measure VPi-ATP in human exercising muscle for the first time. Steady-state VPi-ATP was measured at rest and over a range of exercise intensities and compared with suprabasal oxidative ATP synthesis rates estimated from the initial rates of postexercise phosphocreatine resynthesis (VATP). We define a surplus Pi→ATP flux as the difference between VPi-ATP and VATP. The coupled reactions catalyzed by the glycolytic enzymes GAPDH and phosphoglycerate kinase (PGK) have been shown to catalyze measurable exchange between ATP and Pi in some systems and have been suggested to be responsible for this surplus flux. Surplus VPi-ATP did not change between rest and exercise, even though the concentrations of Pi and ADP, which are substrates for GAPDH and PGK, respectively, increased as expected. However, involvement of these enzymes is suggested by correlations between absolute and surplus Pi→ATP flux, both at rest and during exercise, and the intensity of the phosphomonoester peak in the (31)P NMR spectrum. This peak includes contributions from sugar phosphates in the glycolytic pathway, and changes in its intensity may indicate changes in downstream glycolytic intermediates, including 3-phosphoglycerate, which has been shown to influence the exchange between ATP and Pi catalyzed by GAPDH and PGK. Copyright © 2016 the American Physiological Society.

[ATP content in cryopreserved sperm of Siberian white cranes Grus leucogeranus (Aves: Gruiformes)].

PubMed

Maksudov, G Iu; Erokhin, A S; Nesterenko, O N; Panchenko, V G

2002-01-01

ATP contents were studied in the native and cryoconserved sperm of Siberian white cranes Grus leucogeranus using bioluminescence analysis. The ATP content in freshly obtained spermatozoa was 12.7 nmol/10(8) cells. No ATP was found in the seminal plasma. In the process of freezing-thawing, the ATP concentration in the spermatozoa decreased by 30%. The differences in the dynamics of ATP content during cryoconservation of sperm of white cranes and other birds and mammals are discussed.

Facile conversion of ATP-binding RNA aptamer to quencher-free molecular aptamer beacon.

PubMed

Park, Yoojin; Nim-Anussornkul, Duangrat; Vilaivan, Tirayut; Morii, Takashi; Kim, Byeang Hyean

2018-01-15

We have developed RNA-based quencher-free molecular aptamer beacons (RNA-based QF-MABs) for the detection of ATP, taking advantage of the conformational changes associated with ATP binding to the ATP-binding RNA aptamer. The RNA aptamer, with its well-defined structure, was readily converted to the fluorescence sensors by incorporating a fluorophore into the loop region of the hairpin structure. These RNA-based QF-MABs exhibited fluorescence signals in the presence of ATP relative to their low background signals in the absence of ATP. The fluorescence emission intensity increased upon formation of a RNA-based QF-MAB·ATP complex. Copyright © 2017 Elsevier Ltd. All rights reserved.

Real-time imaging of inflation-induced ATP release in the ex vivo rat lung.

PubMed

Furuya, Kishio; Tan, Ju Jing; Boudreault, Francis; Sokabe, Masahiro; Berthiaume, Yves; Grygorczyk, Ryszard

2016-11-01

Extracellular ATP and other nucleotides are important autocrine/paracrine mediators that regulate diverse processes critical for lung function, including mucociliary clearance, surfactant secretion, and local blood flow. Cellular ATP release is mechanosensitive; however, the impact of physical stimuli on ATP release during breathing has never been tested in intact lungs in real time and remains elusive. In this pilot study, we investigated inflation-induced ATP release in rat lungs ex vivo by real-time luciferin-luciferase (LL) bioluminescence imaging coupled with simultaneous infrared tissue imaging to identify ATP-releasing sites. With LL solution introduced into air spaces, brief inflation of such edematous lung (1 s, ∼20 cmH 2 O) induced transient (<30 s) ATP release in a limited number of air-inflated alveolar sacs during their recruitment/opening. Released ATP reached concentrations of ∼10 -6 M, relevant for autocrine/paracrine signaling, but it remained spatially restricted to single alveolar sacs or their clusters. ATP release was stimulus dependent: prolonged (100 s) inflation evoked long-lasting ATP release that terminated upon alveoli deflation/derecruitment while cyclic inflation/suction produced cyclic ATP release. With LL introduced into blood vessels, inflation induced transient ATP release in many small patchlike areas the size of alveolar sacs. Findings suggest that inflation induces ATP release in both alveoli and the surrounding blood capillary network; the functional units of ATP release presumably consist of alveolar sacs or their clusters. Our study demonstrates the feasibility of real-time ATP release imaging in ex vivo lungs and provides the first direct evidence of inflation-induced ATP release in lung air spaces and in pulmonary blood capillaries, highlighting the importance of purinergic signaling in lung function. Copyright © 2016 the American Physiological Society.

Diadenosine tetraphosphate-gating of cardiac K(ATP) channels requires intact actin cytoskeleton.

PubMed

Jovanović, S; Jovanović, A

2001-09-01

Diadenosine polyphosphates (ApnA) have been recently discovered in the heart, and their levels found to be regulated by ischemia. These signaling molecules are believed to regulate cellular processes that alarm a cell to metabolic stress. In particular, changes in cardiac diadenosine polyphosphates (ApnA) levels may contribute to the regulation of ATP-sensitive K+ (K(ATP)) channel activity, an ion channel that couples the cellular metabolic state with membrane excitability. A feature of myocardial ischemia is the disruption of the actin cytoskeleton which critically regulates the behavior of K(ATP) channels. Whether the integrity of actin microfilaments regulates the interaction of ApnA with K(ATP) channels is not known. The inside-out configuration of the patch-clamp technique was applied to cardiomyocytes isolated from guinea-pig heart. Following patch excision, the prototype dinucleotide, diadenosine tetraphosphate (Ap4A), inhibited K(ATP) channel opening. Treatment of the internal side of membrane patches with either cytochalasin B or DNase I, disrupters of the actin cytoskeleton, prevented Ap4A-induced inhibition of K(ATP) channel opening. Application of purified actin to DNase-treated membrane patches restored the ability of Ap4A to close K(ATP) channels. This study shows that inhibition of cardiac K(ATP) channel by Ap4A, a putative alarmone, requires intact subsarcolemmal actin network. Such interaction between K(ATP) channels, the cardiomyocyte cytoskeleton and intracellular Ap4A could affect different channel-dependent functions.

ATP Synthase: A Molecular Therapeutic Drug Target for Antimicrobial and Antitumor Peptides

PubMed Central

Ahmad, Zulfiqar; Okafor, Florence; Azim, Sofiya; Laughlin, Thomas F.

2015-01-01

In this review we discuss the role of ATP synthase as a molecular drug target for natural and synthetic antimi-crobial/antitumor peptides. We start with an introduction of the universal nature of the ATP synthase enzyme and its role as a biological nanomotor. Significant structural features required for catalytic activity and motor functions of ATP synthase are described. Relevant details regarding the presence of ATP synthase on the surface of several animal cell types, where it is associated with multiple cellular processes making it a potential drug target with respect to antimicrobial peptides and other inhibitors such as dietary polyphenols, is also reviewed. ATP synthase is known to have about twelve discrete inhibitor binding sites including peptides and other inhibitors located at the interface of α/β subunits on the F1 sector of the enzyme. Molecular interaction of peptides at the β DEELSEED site on ATP synthase is discussed with specific examples. An inhibitory effect of other natural/synthetic inhibitors on ATP is highlighted to explore the therapeutic roles played by peptides and other inhibitors. Lastly, the effect of peptides on the inhibition of the Escherichia coli model system through their action on ATP synthase is presented. PMID:23432591

[Crystallography of ATP hydrolysis mechanism in rat brain kinesin].

PubMed

Wan, Qun; Zhu, Pingting; Lü, Houning; Chen, Xinhong

2014-04-01

Rat brain kinesin is a conventional kinesin that uses the energy from ATP hydrolysis to walk along the microtubule progressively. Studying how the chemical energy in ATP is utilized for mechanical movement is important to understand this moving function. The monomeric motor domain, rK354, was crystallized. An ATP analog, AMPPNP, was soaked in the active site. Comparing the complex structure of rK354 x AMPPNP and that of rK354ADP, a hypothesis is proposed that Glu237 in the Switch II region sensors the presence of gamma-phosphate and transfers the signal to the microtubule binding region.

How Native and Alien Metal Cations Bind ATP: Implications for Lithium as a Therapeutic Agent

NASA Astrophysics Data System (ADS)

Dudev, Todor; Grauffel, Cédric; Lim, Carmay

2017-02-01

Adenosine triphosphate (ATP), the major energy currency of the cell, exists in solution mostly as ATP-Mg. Recent experiments suggest that Mg2+ interacts with the highly charged ATP triphosphate group and Li+ can co-bind with the native Mg2+ to form ATP-Mg-Li and modulate the neuronal purine receptor response. However, it is unclear how the negatively charged ATP triphosphate group binds Mg2+ and Li+ (i.e. which phosphate group(s) bind Mg2+/Li+) and how the ATP solution conformation depends on the type of metal cation and the metal-binding mode. Here, we reveal the preferred ATP-binding mode of Mg2+/Li+ alone and combined: Mg2+ prefers to bind ATP tridentately to each of the three phosphate groups, but Li+ prefers to bind bidentately to the terminal two phosphates. We show that the solution ATP conformation depends on the cation and its binding site/mode, but it does not change significantly when Li+ binds to Mg2+-loaded ATP. Hence, ATP-Mg-Li, like Mg2+-ATP, can fit in the ATP-binding site of the host enzyme/receptor, activating specific signaling pathways.

A futile cycle, formed between two ATP-dependant gamma-glutamyl cycle enzymes, gamma-glutamyl cysteine synthetase and 5-oxoprolinase: the cause of cellular ATP depletion in nephrotic cystinosis?

PubMed

Kumar, Akhilesh; Bachhawat, Anand Kumar

2010-03-01

Cystinosis, an inherited disease caused by a defect in the lysosomal cystine transporter (CTNS), is characterized by renal proximal tubular dysfunction. Adenosine triphosphate (ATP) depletion appears to be a key event in the pathophysiology of the disease, even though the manner in which ATP depletion occurs is still a puzzle. We present a model that explains how a futile cycle that is generated between two ATP-utilizing enzymes of the gamma-glutamyl cycle leads to ATP depletion. The enzyme gamma-glutamyl cysteine synthetase (gamma-GCS), in the absence of cysteine, forms 5-oxoproline (instead of the normal substrate, gamma-glutamyl cysteine) and the 5-oxoproline is converted into glutamate by the ATP-dependant enzyme, 5-oxoprolinase. Thus, in cysteine-limiting conditions, glutamate is cycled back into glutamate via 5-oxoproline at the cost of two ATP molecules without production of glutathione and is the cause of the decreased levels of glutathione synthesis, as well as the ATP depletion observed in these cells. The model is also compatible with the differences seen in the human patients and the mouse model of cystinosis, where renal failure is not observed.

Global gene profiling of aging lungs in Atp8b1 mutant mice.

PubMed

Soundararajan, Ramani; Stearns, Timothy M; Czachor, Alexander; Fukumoto, Jutaro; Turn, Christina; Westermann-Clark, Emma; Breitzig, Mason; Tan, Lee; Lockey, Richard F; King, Benjamin L; Kolliputi, Narasaiah

2016-09-29

Recent studies implicate cardiolipin oxidation in several age-related diseases. Atp8b1 encoding Type 4 P-type ATPases is a cardiolipin transporter. Mutation in Atp8b1 gene or inflammation of the lungs impairs the capacity of Atp8b1 to clear cardiolipin from lung fluid. However, the link between Atp8b1 mutation and age-related gene alteration is unknown. Therefore, we investigated how Atp8b1 mutation alters age-related genes. We performed Affymetrix gene profiling of lungs isolated from young (7-9 wks, n=6) and aged (14 months, 14 M, n=6) C57BL/6 and Atp8b1 mutant mice. In addition, Ingenuity Pathway Analysis (IPA) was performed. Differentially expressed genes were validated by quantitative real-time PCR (qRT-PCR). Global transcriptome analysis revealed 532 differentially expressed genes in Atp8b1 lungs, 157 differentially expressed genes in C57BL/6 lungs, and 37 overlapping genes. IPA of age-related genes in Atp8b1 lungs showed enrichment of Xenobiotic metabolism and Nrf2-mediated signaling pathways. The increase in Adamts2 and Mmp13 transcripts in aged Atp8b1 lungs was validated by qRT-PCR. Similarly, the decrease in Col1a1 and increase in Cxcr6 transcripts was confirmed in both Atp8b1 mutant and C57BL/6 lungs. Based on transcriptome profiling, our study indicates that Atp8b1 mutant mice may be susceptible to age-related lung diseases.

General anesthetics cause mitochondrial dysfunction and reduction of intracellular ATP levels

PubMed Central

Kishikawa, Jun-ichi; Inoue, Yuki; Fujikawa, Makoto; Nishimura, Kenji; Nakanishi, Atsuko; Tanabe, Tsutomu; Imamura, Hiromi

2018-01-01

General anesthetics are indispensable for effective clinical care. Although, the mechanism of action of general anesthetics remains controversial, lipid bilayers and proteins have been discussed as their targets. In this study, we focused on the relationship between cellular ATP levels and general anesthetics. The ATP levels of nematodes and cultured mammalian cells were decreased by exposure to three general anesthetics: isoflurane, pentobarbital, and 1-phenoxy-2-propanol. Furthermore, these general anesthetics abolished mitochondrial membrane potential, resulting in the inhibition of mitochondrial ATP synthesis. These results suggest that the observed decrease of cellular ATP level is a common phenomenon of general anesthetics. PMID:29298324

Determination of Turboprop Reduction Gearbox System Fatigue Life and Reliability

NASA Technical Reports Server (NTRS)

Zaretsky, Erwin V.; Lewicki, David G.; Savage, Michael; Vlcek, Brian L.

2007-01-01

Two computational models to determine the fatigue life and reliability of a commercial turboprop gearbox are compared with each other and with field data. These models are (1) Monte Carlo simulation of randomly selected lives of individual bearings and gears comprising the system and (2) two-parameter Weibull distribution function for bearings and gears comprising the system using strict-series system reliability to combine the calculated individual component lives in the gearbox. The Monte Carlo simulation included the virtual testing of 744,450 gearboxes. Two sets of field data were obtained from 64 gearboxes that were first-run to removal for cause, were refurbished and placed back in service, and then were second-run until removal for cause. A series of equations were empirically developed from the Monte Carlo simulation to determine the statistical variation in predicted life and Weibull slope as a function of the number of gearboxes failed. The resultant L(sub 10) life from the field data was 5,627 hr. From strict-series system reliability, the predicted L(sub 10) life was 774 hr. From the Monte Carlo simulation, the median value for the L(sub 10) gearbox lives equaled 757 hr. Half of the gearbox L(sub 10) lives will be less than this value and the other half more. The resultant L(sub 10) life of the second-run (refurbished) gearboxes was 1,334 hr. The apparent load-life exponent p for the roller bearings is 5.2. Were the bearing lives to be recalculated with a load-life exponent p equal to 5.2, the predicted L(sub 10) life of the gearbox would be equal to the actual life obtained in the field. The component failure distribution of the gearbox from the Monte Carlo simulation was nearly identical to that using the strict-series system reliability analysis, proving the compatibility of these methods.

An uracil-linked hydroxyflavone probe for the recognition of ATP

PubMed Central

Bojtár, Márton; Janzsó-Berend, Péter Zoltán; Mester, Dávid; Hessz, Dóra; Kállay, Mihály; Kubinyi, Miklós

2018-01-01

Background: Nucleotides are essential molecules in living systems due to their paramount importance in various physiological processes. In the past years, numerous attempts were made to selectively recognize and detect these analytes, especially ATP using small-molecule fluorescent chemosensors. Despite the various solutions, the selective detection of ATP is still challenging due to the structural similarity of various nucleotides. In this paper, we report the conjugation of a uracil nucleobase to the known 4’-dimethylamino-hydroxyflavone fluorophore. Results: The complexation of this scaffold with ATP is already known. The complex is held together by stacking and electrostatic interactions. To achieve multi-point recognition, we designed the uracil-appended version of this probe to include complementary base-pairing interactions. The theoretical calculations revealed the availability of multiple complex structures. The synthesis was performed using click chemistry and the nucleotide recognition properties of the probe were evaluated using fluorescence spectroscopy. Conclusions: The first, uracil-containing fluorescent ATP probe based on a hydroxyflavone fluorophore was synthesized and evaluated. A selective complexation with ATP was observed and a ratiometric response in the excitation spectrum. PMID:29719572

Developing and utilizing an Euler computational method for predicting the airframe/propulsion effects for an aft-mounted turboprop transport. Volume 2: User guide

NASA Technical Reports Server (NTRS)

Chen, H. C.; Neback, H. E.; Kao, T. J.; Yu, N. Y.; Kusunose, K.

1991-01-01

This manual explains how to use an Euler based computational method for predicting the airframe/propulsion integration effects for an aft-mounted turboprop transport. The propeller power effects are simulated by the actuator disk concept. This method consists of global flow field analysis and the embedded flow solution for predicting the detailed flow characteristics in the local vicinity of an aft-mounted propfan engine. The computational procedure includes the use of several computer programs performing four main functions: grid generation, Euler solution, grid embedding, and streamline tracing. This user's guide provides information for these programs, including input data preparations with sample input decks, output descriptions, and sample Unix scripts for program execution in the UNICOS environment.

[Change in concentration of cytosolic Ca2+ caused by extracellular ATP and ecto-ATP-ase activity in thymocytes and transformed MT-4 cells].

PubMed

Hrebinyk, S M; Artemenko, O Iu; Hryniuk, I I; Perepelitsyna, O M; Matyshevs'ka, O P

2009-01-01

The comparative study of extracellular ATP (ATP0) effect on free cytosolic calcium concentration ([Ca2+]i) in normal (isolated rat thymocytes) and transformed (leukosis MT-4 line) T-cells was carried out. Addition of 1 mM ATP to Ca-free incubation medium of both types of cells, loaded with indo-1, had no effect on [Ca2+]i level. Upon subsequent addition of 1 mM CaCl2 to the incubation medium the rapid and significant increase of [Ca2+]i in MT-4 cells was registered. This effect was maintained within 10 min and was not inhibited by phospholipase C inhibitor 0.2 mM neomycin, that was induced by cation entry into the cells from the extracellular medium. Both types of cells were shown to demonstrate ecto-ATPase activity in the presence of 1 mM MgCl2 or CaC12 in the incubation medium. Estimation of kinetic parameters has indicated that the maximum rate of extracellular ATP hydrolysis by MT-4 cells is higher and Mg2+ and Ca2+ activation constants are lower as compared to respective parameters of ATP hydrolysis by thymocytes. The possible functional significance of the increased level of ecto-ATPase activity in malignantly transformed cells is discussed.

ATP7B mediates vesicular sequestration of copper: insight into biliary copper excretion.

PubMed

Cater, Michael A; La Fontaine, Sharon; Shield, Kristy; Deal, Yolanda; Mercer, Julian F B

2006-02-01

The Wilson protein (ATP7B) regulates levels of systemic copper by excreting excess copper into bile. It is not clear whether ATP7B translocates excess intrahepatic copper directly across the canalicular membrane or sequesters this copper into exocytic vesicles, which subsequently fuse with canalicular membrane to expel their contents into bile. The aim of this study was to clarify the mechanism underlying ATP7B-mediated copper detoxification by investigating endogenous ATP7B localization in the HepG2 hepatoma cell line and its ability to mediate vesicular sequestration of excess intracellular copper. Immunofluorescence microscopy was used to investigate the effect of copper concentration on the localization of endogenous ATP7B in HepG2 cells. Copper accumulation studies to determine whether ATP7B can mediate vesicular sequestration of excess intracellular copper were performed using Chinese hamster ovary cells that exogenously expressed wild-type and mutant ATP7B proteins. In HepG2 cells, elevated copper levels stimulated trafficking of ATP7B to pericanalicular vesicles and not to the canalicular membrane as previously reported. Mutation of an endocytic retrieval signal in ATP7B caused the protein to constitutively localize to vesicles and not to the plasma membrane, suggesting that a vesicular compartment(s) is the final trafficking destination for ATP7B. Expression of wild-type and mutant ATP7B caused Chinese hamster ovary cells to accumulate copper in vesicles, which subsequently undergo exocytosis, releasing copper across the plasma membrane. This report provides compelling evidence that the primary mechanism of biliary copper excretion involves ATP7B-mediated vesicular sequestration of copper rather than direct copper translocation across the canalicular membrane.

CFD analysis of turboprop engine oil cooler duct for best rate of climb condition

NASA Astrophysics Data System (ADS)

Kalia, Saurabh; CA, Vinay; Hegde, Suresh M.

2016-09-01

Turboprop engines are widely used in commuter category airplanes. Aircraft Design bureaus routinely conduct the flight tests to confirm the performance of the system. The lubrication system of the engine is designed to provide a constant supply of clean lubrication oil to the engine bearings, the reduction gears, the torque-meter, the propeller and the accessory gearbox. The oil lubricates, cools and also conducts foreign material to the oil filter where it is removed from further circulation. Thus a means of cooling the engine oil must be provided and a suitable oil cooler (OC) and ducting system was selected and designed for this purpose. In this context, it is relevant to study and analyse behaviour of the engine oil cooler system before commencing actual flight tests. In this paper, the performance of the oil cooler duct with twin flush NACA inlet housed inside the nacelle has been studied for aircraft best rate of climb (ROC) condition using RANS based SST K-omega model by commercial software ANSYS Fluent 13.0. From the CFD analysis results, it is found that the mass flow rate captured and pressure drop across the oil cooler for the best ROC condition is meeting the oil cooler manufacturer requirements thus, the engine oil temperature is maintained within prescribed limits.

PSHFT - COMPUTERIZED LIFE AND RELIABILITY MODELLING FOR TURBOPROP TRANSMISSIONS

NASA Technical Reports Server (NTRS)

Savage, M.

1994-01-01

The computer program PSHFT calculates the life of a variety of aircraft transmissions. A generalized life and reliability model is presented for turboprop and parallel shaft geared prop-fan aircraft transmissions. The transmission life and reliability model is a combination of the individual reliability models for all the bearings and gears in the main load paths. The bearing and gear reliability models are based on the statistical two parameter Weibull failure distribution method and classical fatigue theories. The computer program developed to calculate the transmission model is modular. In its present form, the program can analyze five different transmissions arrangements. Moreover, the program can be easily modified to include additional transmission arrangements. PSHFT uses the properties of a common block two-dimensional array to separate the component and transmission property values from the analysis subroutines. The rows correspond to specific components with the first row containing the values for the entire transmission. Columns contain the values for specific properties. Since the subroutines (which determine the transmission life and dynamic capacity) interface solely with this property array, they are separated from any specific transmission configuration. The system analysis subroutines work in an identical manner for all transmission configurations considered. Thus, other configurations can be added to the program by simply adding component property determination subroutines. PSHFT consists of a main program, a series of configuration specific subroutines, generic component property analysis subroutines, systems analysis subroutines, and a common block. The main program selects the routines to be used in the analysis and sequences their operation. The series of configuration specific subroutines input the configuration data, perform the component force and life analyses (with the help of the generic component property analysis subroutines), fill the

The scaffold protein calcium/calmodulin-dependent serine protein kinase controls ATP release in sensory ganglia upon P2X3 receptor activation and is part of an ATP keeper complex.

PubMed

Bele, Tanja; Fabbretti, Elsa

2016-08-01

P2X3 receptors, gated by extracellular ATP, are expressed by sensory neurons and are involved in peripheral nociception and pain sensitization. The ability of P2X3 receptors to transduce extracellular stimuli into neuronal signals critically depends on the dynamic molecular partnership with the calcium/calmodulin-dependent serine protein kinase (CASK). The present work used trigeminal sensory neurons to study the impact that activation of P2X3 receptors (evoked by the agonist α,β-meATP) has on the release of endogenous ATP and how CASK modulates this phenomenon. P2X3 receptor function was followed by ATP efflux via Pannexin1 (Panx1) hemichannels, a mechanism that was blocked by the P2X3 receptor antagonist A-317491, and by P2X3 silencing. ATP efflux was enhanced by nerve growth factor, a treatment known to potentiate P2X3 receptor function. Basal ATP efflux was not controlled by CASK, and carbenoxolone or Pannexin silencing reduced ATP release upon P2X3 receptor function. CASK-controlled ATP efflux followed P2X3 receptor activity, but not depolarization-evoked ATP release. Molecular biology experiments showed that CASK was essential for the transactivation of Panx1 upon P2X3 receptor activation. These data suggest that P2X3 receptor function controls a new type of feed-forward purinergic signaling on surrounding cells, with consequences at peripheral and spinal cord level. Thus, P2X3 receptor-mediated ATP efflux may be considered for the future development of pharmacological strategies aimed at containing neuronal sensitization. P2X3 receptors are involved in sensory transduction and associate to CASK. We have studied in primary sensory neurons the molecular mechanisms downstream P2X3 receptor activation, namely ATP release and partnership with CASK or Panx1. Our data suggest that CASK and P2X3 receptors are part of an ATP keeper complex, with important feed-forward consequences at peripheral and central level. © 2016 International Society for Neurochemistry.

Lipidomic and metabolic changes in the P4-type ATPase ATP10D deficient C57BL/6J wild type mice upon rescue of ATP10D function.

PubMed

Sigruener, Alexander; Wolfrum, Christian; Boettcher, Alfred; Kopf, Thomas; Liebisch, Gerhard; Orsó, Evelyn; Schmitz, Gerd

2017-01-01

Sequence variants near the human gene for P4-type ATPase, class V, type 10D (ATP10D) were shown to significantly associate with circulating hexosylceramide d18:1/16:0 and d18:1/24:1 levels, obesity, insulin resistance, plasma high density lipoprotein (HDL), coronary stenotic index and intracranial atherosclerotic index. In mice Atp10d is associated with HDL modulation and C57BL/6 mice expressing a truncated, non-functional form of ATP10D easily develop obesity and insulin resistance on high-fat diet. We analyzed metabolic differences of ATP10D deficient C57BL/6J wild type and ATP10D transgenic C57BL/6J BAC129 mice. ATP10D transgenic mice gain 25% less weight on high-fat diet concomitant with a reduced increase in fat cell mass but independent of adipocyte size change. ATP10D transgenic mice also had 26% lower triacylglycerol levels with approximately 76% bound to very low density lipoprotein while in ATP10D deficient wild type mice 57% are bound to low density lipoprotein. Furthermore increased oxygen consumption and CO2 production, 38% lower glucose and 69% lower insulin levels and better insulin sensitivity were observed in ATP10D transgenic mice. Besides decreased hexosylceramide species levels were detected. Part of these effects may be due to reduced hepatic stearoyl-CoA desaturase 1 (SCD1) expression in ATP10D transgenic mice, which was reflected by altered fatty acid and lipid species patterns. There was a significant decrease in the hepatic 18:1 to 18:0 free fatty acid ratio in transgenic mice. The ratio of 16:1 to 16:0 was not significantly different. Interestingly both ratios were significantly reduced in plasma total fatty acids. In summary we found that ATP10D reduces high-fat diet induced obesity and improves insulin sensitivity. ATP10D transgenic mice showed altered hepatic expression of lipid-metabolism associated genes, including Scd1, along with changes in hepatic and plasma lipid species and plasma lipoprotein pattern.

Synergistic binding of glucose and aluminium ATP to hexokinase from Saccharomyces cerevisiae.

PubMed

Woolfitt, A R; Kellett, G L; Hoggett, J G

1988-08-10

The binding of glucose, AlATP and AlADP to the monomeric and dimeric forms of the native yeast hexokinase PII isoenzyme and to the proteolytically modified SII monomeric form was monitored at pH 6.7 by the concomitant quenching of intrinsic protein fluorescence. No fluorescence changes were observed when free enzyme was mixed with AlATP at concentrations up to 7500 microM. In the presence of saturating concentrations of glucose, the maximal quenching of fluorescence induced by AlATP was between 1.5 and 3.5% depending on species, and the average value of [L]0.5, the concentration of ligand at half-saturation, over all monomeric species was 0.9 +/- 0.4 microM. The presence of saturating concentrations of AlATP diminished [L]0.5 for glucose binding by between 260- and 670-fold for hexokinase PII and SII monomers, respectively (dependent on the ionic strength), and by almost 4000-fold for PII dimer. The data demonstrate extremely strong synergistic interactions in the binding of glucose and AlATP to yeast hexokinase, arising as a consequence of conformational changes in the free enzyme induced by glucose and in enzyme-glucose complex induced by AlATP. The synergistic interactions of glucose and AlATP are related to their kinetic synergism and to the ability of AlATP to act as a powerful inhibitor of the hexokinase reaction.

Regulation of calreticulin–major histocompatibility complex (MHC) class I interactions by ATP

PubMed Central

Wijeyesakere, Sanjeeva Joseph; Gagnon, Jessica K.; Arora, Karunesh; Brooks, Charles L.; Raghavan, Malini

2015-01-01

The MHC class I peptide loading complex (PLC) facilitates the assembly of MHC class I molecules with peptides, but factors that regulate the stability and dynamics of the assembly complex are largely uncharacterized. Based on initial findings that ATP, in addition to MHC class I-specific peptide, is able to induce MHC class I dissociation from the PLC, we investigated the interaction of ATP with the chaperone calreticulin, an endoplasmic reticulum (ER) luminal, calcium-binding component of the PLC that is known to bind ATP. We combined computational and experimental measurements to identify residues within the globular domain of calreticulin, in proximity to the high-affinity calcium-binding site, that are important for high-affinity ATP binding and for ATPase activity. High-affinity calcium binding by calreticulin is required for optimal nucleotide binding, but both ATP and ADP destabilize enthalpy-driven high-affinity calcium binding to calreticulin. ATP also selectively destabilizes the interaction of calreticulin with cellular substrates, including MHC class I molecules. Calreticulin mutants that affect ATP or high-affinity calcium binding display prolonged associations with monoglucosylated forms of cellular MHC class I, delaying MHC class I dissociation from the PLC and their transit through the secretory pathway. These studies reveal central roles for ATP and calcium binding as regulators of calreticulin–substrate interactions and as key determinants of PLC dynamics. PMID:26420867

A lipid switch unlocks Parkinson’s disease-associated ATP13A2

PubMed Central

Holemans, Tine; Sørensen, Danny Mollerup; van Veen, Sarah; Martin, Shaun; Hermans, Diane; Kemmer, Gerdi Christine; Van den Haute, Chris; Baekelandt, Veerle; Günther Pomorski, Thomas; Agostinis, Patrizia; Wuytack, Frank; Palmgren, Michael; Eggermont, Jan; Vangheluwe, Peter

2015-01-01

ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor–Rakeb syndrome and Parkinson’s disease (PD), providing protection against α-synuclein, Mn2+, and Zn2+ toxicity in various model systems. So far, the molecular function and regulation of ATP13A2 remains undetermined. Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. PMID:26134396

Effects and mechanism of acid rain on plant chloroplast ATP synthase.

PubMed

Sun, Jingwen; Hu, Huiqing; Li, Yueli; Wang, Lihong; Zhou, Qing; Huang, Xiaohua

2016-09-01

Acid rain can directly or indirectly affect plant physiological functions, especially photosynthesis. The enzyme ATP synthase is the key in photosynthetic energy conversion, and thus, it affects plant photosynthesis. To clarify the mechanism by which acid rain affects photosynthesis, we studied the effects of acid rain on plant growth, photosynthesis, chloroplast ATP synthase activity and gene expression, chloroplast ultrastructure, intracellular H(+) level, and water content of rice seedlings. Acid rain at pH 4.5 remained the chloroplast structure unchanged but increased the expression of six chloroplast ATP synthase subunits, promoted chloroplast ATP synthase activity, and increased photosynthesis and plant growth. Acid rain at pH 4.0 or less decreased leaf water content, destroyed chloroplast structure, inhibited the expression of six chloroplast ATP synthase subunits, decreased chloroplast ATP synthase activity, and reduced photosynthesis and plant growth. In conclusion, acid rain affected the chloroplast ultrastructure, chloroplast ATPase transcription and activity, and P n by changing the acidity in the cells, and thus influencing the plant growth and development. Finally, the effects of simulated acid rain on the test indices were found to be dose-dependent.

Determination of effects of ambient conditions on aircraft engine emissions engine testing - GTCP 85 APU, TPE 331 turboprop. Volume 2. Final report, 20 Dec 1974--20 Dec 1975

DOE Office of Scientific and Technical Information (OSTI.GOV)

Slogar, G.A.; Holder, R.C.

1976-03-01

Full scale engine testswere conducted on a GTCP85-98CK Auxiliary Power Unit and a TPE331-5-251M Turboprop engine. The purpose of this program was to measure exhaust emission of HC, CO, CO/sub 2/, NO/sub x/, and smoke at controlled (temperature, humidity, and pressure) engine inlet conditions. This data along with other available data will provide the data base for the determination of the effects of ambient conditions on gas turbine engines. This volume contains the computer programs for volume 2 data. (GRA)

ATP synthase--a marvellous rotary engine of the cell.

PubMed

Yoshida, M; Muneyuki, E; Hisabori, T

2001-09-01

ATP synthase can be thought of as a complex of two motors--the ATP-driven F1 motor and the proton-driven Fo motor--that rotate in opposite directions. The mechanisms by which rotation and catalysis are coupled in the working enzyme are now being unravelled on a molecular scale.

Light-driven production of ATP catalysed by F0F1-ATP synthase in an artificial photosynthetic membrane

NASA Astrophysics Data System (ADS)

Steinberg-Yfrach, Gali; Rigaud, Jean-Louis; Durantini, Edgardo N.; Moore, Ana L.; Gust, Devens; Moore, Thomas A.

1998-04-01

Energy-transducing membranes of living organisms couple spontaneous to non-spontaneous processes through the intermediacy of protonmotive force (p.m.f.) - an imbalance in electrochemical potential of protons across the membrane. In most organisms, p.m.f. is generated by redox reactions that are either photochemically driven, such as those in photosynthetic reaction centres, or intrinsically spontaneous, such as those of oxidative phosphorylation in mitochondria. Transmembrane proteins (such as the cytochromes and complexes I, III and IV in the electron-transport chain in the inner mitochondrial membrane) couple the redox reactions to proton translocation, thereby conserving a fraction of the redox chemical potential as p.m.f. Many transducer proteins couple p.m.f. to the performance of biochemical work, such as biochemical synthesis and mechanical and transport processes. Recently, an artificial photosynthetic membrane was reported in which a photocyclic process was used to transport protons across a liposomal membrane, resulting in acidification of the liposome's internal volume. If significant p.m.f. is generated in this system, then incorporating an appropriate transducer into the liposomal bilayer should make it possible to drive a non-spontaneous chemical process. Here we report the incorporation of FOF1-ATP synthase into liposomes containing the components of the proton-pumping photocycle. Irradiation of this artificial membrane with visible light results in the uncoupler- and inhibitor-sensitive synthesis of adenosine triphosphate (ATP) against an ATP chemical potential of ~12kcalmol-1, with a quantum yield of more than 7%. This system mimics the process by which photosynthetic bacteria convert light energy into ATP chemical potential.

Origin Licensing Requires ATP Binding and Hydrolysis by the MCM Replicative Helicase

PubMed Central

Coster, Gideon; Frigola, Jordi; Beuron, Fabienne; Morris, Edward P.; Diffley, John F.X.

2014-01-01

Summary Loading of the six related Minichromosome Maintenance (MCM) proteins as head-to-head double hexamers during DNA replication origin licensing is crucial for ensuring once-per-cell-cycle DNA replication in eukaryotic cells. Assembly of these prereplicative complexes (pre-RCs) requires the Origin Recognition Complex (ORC), Cdc6, and Cdt1. ORC, Cdc6, and MCM are members of the AAA+ family of ATPases, and pre-RC assembly requires ATP hydrolysis. Here we show that ORC and Cdc6 mutants defective in ATP hydrolysis are competent for origin licensing. However, ATP hydrolysis by Cdc6 is required to release nonproductive licensing intermediates. We show that ATP binding stabilizes the wild-type MCM hexamer. Moreover, by analyzing MCM containing mutant subunits, we show that ATP binding and hydrolysis by MCM are required for Cdt1 release and double hexamer formation. This work alters our view of how ATP is used by licensing factors to assemble pre-RCs. PMID:25087873

[Stabilization of Cadmium Contaminated Soils by Ferric Ion Modified Attapulgite (Fe/ATP)--Characterizations and Stabilization Mechanism].

PubMed

Rong, Yang; Li, Rong-bo; Zhou, Yong-li; Chen, Jing; Wang, Lin-ling; Lu, Xiao-hua

2015-08-01

Ferric ion modified attapulgite (Fe/ATP) was prepared by impregnation and its structure and morphology were characterized. The toxicity characteristic leaching procedure (TCLP) was used to evaluate the effect of Cadmium( Cd) stabilization in soil with the addition of attapulgite (ATP) and Fe/ATP. The stabilization mechanism of Cd was further elucidated by comparing the morphologies and structure of ATP and Fe/ATP before and after Cd adsorption. Fe/ATP exhibited much better adsorption capacity than ATP, suggesting different adsorption mechanisms occurred between ATP and Fe/ATP. The leaching concentrations of Cd in soil decreased by 45% and 91% respectively, with the addition of wt. 20% ATP and Fe/ATP. The former was attributed to the interaction between Cd2 and --OH groups by chemical binding to form inner-sphere complexes in ATP and the attachment between Cd2+ and the defect sites in ATP framework. Whereas Cd stabilization with Fe/ATP was resulted from the fact that the active centers (--OH bonds or O- sites) on ATP could react with Fe3+ giving Fe--O--Cd-- bridges, which helped stabilize Cd in surface soil. What'more, the ferric oxides and metal hydroxides on the surface of ATP could interact with Cd, probably by the formation of cadmium ferrite. In conclusion, Fe/ATP, which can be easily prepared, holds promise as a potential low-cost and environmental friendly stabilizing agent for remediation of soil contaminated with heavy metals.

ATP oscillations mediate inductive action of FGF and Shh signalling on prechondrogenic condensation.

PubMed

Kwon, Hyuck Joon

2013-01-01

Skeletal patterns are prefigured by prechondrogenic condensation. Morphogens such as fibroblast growth factor (FGF) and sonic hedgehog (Shh) specify the skeletal patterns in limb development. However, how morphogens regulate prechondrogenic condensation has remained unclear. Recently, it was demonstrated that synchronized Adenosine triphosphate (ATP) oscillations play a critical role in prechondrogenic condensation. Thus, the present study has focused on whether ATP oscillations mediate the actions of major developmental morphogens such as FGF and Shh on prechondrogenic condensation. It has been shown that both FGF and Shh signalling promoted cellular condensation but not chondrogenic differentiation and also induced ATP oscillations. In addition, blockage of FGF and Shh signalling prevented both ATP oscillations and prechondrogenic condensation. Furthermore, it was found that inhibition of ATP oscillations suppressed FGF/Shh-induced prechondrogenic condensation. These results indicate that ATP oscillations mediate the actions of FGF and Shh signalling on prechondrogenic condensation. This study proposes that morphogens organize skeletal patterns via ATP oscillations. Copyright © 2012 John Wiley & Sons, Ltd.

Structure and conformational states of the bovine mitochondrial ATP synthase by cryo-EM.

PubMed

Zhou, Anna; Rohou, Alexis; Schep, Daniel G; Bason, John V; Montgomery, Martin G; Walker, John E; Grigorieff, Nikolaus; Rubinstein, John L

2015-10-06

Adenosine triphosphate (ATP), the chemical energy currency of biology, is synthesized in eukaryotic cells primarily by the mitochondrial ATP synthase. ATP synthases operate by a rotary catalytic mechanism where proton translocation through the membrane-inserted FO region is coupled to ATP synthesis in the catalytic F1 region via rotation of a central rotor subcomplex. We report here single particle electron cryomicroscopy (cryo-EM) analysis of the bovine mitochondrial ATP synthase. Combining cryo-EM data with bioinformatic analysis allowed us to determine the fold of the a subunit, suggesting a proton translocation path through the FO region that involves both the a and b subunits. 3D classification of images revealed seven distinct states of the enzyme that show different modes of bending and twisting in the intact ATP synthase. Rotational fluctuations of the c8-ring within the FO region support a Brownian ratchet mechanism for proton-translocation-driven rotation in ATP synthases.

Structure and conformational states of the bovine mitochondrial ATP synthase by cryo-EM

PubMed Central

Zhou, Anna; Rohou, Alexis; Schep, Daniel G; Bason, John V; Montgomery, Martin G; Walker, John E; Grigorieff, Nikolaus; Rubinstein, John L

2015-01-01

Adenosine triphosphate (ATP), the chemical energy currency of biology, is synthesized in eukaryotic cells primarily by the mitochondrial ATP synthase. ATP synthases operate by a rotary catalytic mechanism where proton translocation through the membrane-inserted FO region is coupled to ATP synthesis in the catalytic F1 region via rotation of a central rotor subcomplex. We report here single particle electron cryomicroscopy (cryo-EM) analysis of the bovine mitochondrial ATP synthase. Combining cryo-EM data with bioinformatic analysis allowed us to determine the fold of the a subunit, suggesting a proton translocation path through the FO region that involves both the a and b subunits. 3D classification of images revealed seven distinct states of the enzyme that show different modes of bending and twisting in the intact ATP synthase. Rotational fluctuations of the c8-ring within the FO region support a Brownian ratchet mechanism for proton-translocation-driven rotation in ATP synthases. DOI: http://dx.doi.org/10.7554/eLife.10180.001 PMID:26439008

Characterization of the Saccharomyces cerevisiae ATP-Interactome using the iTRAQ-SPROX Technique

NASA Astrophysics Data System (ADS)

Geer, M. Ariel; Fitzgerald, Michael C.

2016-02-01

The stability of proteins from rates of oxidation (SPROX) technique was used in combination with an isobaric mass tagging strategy to identify adenosine triphosphate (ATP) interacting proteins in the Saccharomyces cerevisiae proteome. The SPROX methodology utilized in this work enabled 373 proteins in a yeast cell lysate to be assayed for ATP interactions (both direct and indirect) using the non-hydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP). A total of 28 proteins were identified with AMP-PNP-induced thermodynamic stability changes. These protein hits included 14 proteins that were previously annotated as ATP-binding proteins in the Saccharomyces Genome Database (SGD). The 14 non-annotated ATP-binding proteins included nine proteins that were previously found to be ATP-sensitive in an earlier SPROX study using a stable isotope labeling with amino acids in cell culture (SILAC)-based approach. A bioinformatics analysis of the protein hits identified here and in the earlier SILAC-SPROX experiments revealed that many of the previously annotated ATP-binding protein hits were kinases, ligases, and chaperones. In contrast, many of the newly discovered ATP-sensitive proteins were not from these protein classes, but rather were hydrolases, oxidoreductases, and nucleic acid-binding proteins.

Quantifying intracellular rates of glycolytic and oxidative ATP production and consumption using extracellular flux measurements

PubMed Central

Mookerjee, Shona A.; Gerencser, Akos A.; Nicholls, David G.; Brand, Martin D.

2017-01-01

Partitioning of ATP generation between glycolysis and oxidative phosphorylation is central to cellular bioenergetics but cumbersome to measure. We describe here how rates of ATP generation by each pathway can be calculated from simultaneous measurements of extracellular acidification and oxygen consumption. We update theoretical maximum ATP yields by mitochondria and cells catabolizing different substrates. Mitochondrial P/O ratios (mol of ATP generated per mol of [O] consumed) are 2.73 for oxidation of pyruvate plus malate and 1.64 for oxidation of succinate. Complete oxidation of glucose by cells yields up to 33.45 ATP/glucose with a maximum P/O of 2.79. We introduce novel indices to quantify bioenergetic phenotypes. The glycolytic index reports the proportion of ATP production from glycolysis and identifies cells as primarily glycolytic (glycolytic index > 50%) or primarily oxidative. The Warburg effect is a chronic increase in glycolytic index, quantified by the Warburg index. Additional indices quantify the acute flexibility of ATP supply. The Crabtree index and Pasteur index quantify the responses of oxidative and glycolytic ATP production to alterations in glycolysis and oxidative reactions, respectively; the supply flexibility index quantifies overall flexibility of ATP supply; and the bioenergetic capacity quantifies the maximum rate of total ATP production. We illustrate the determination of these indices using C2C12 myoblasts. Measurement of ATP use revealed no significant preference for glycolytic or oxidative ATP by specific ATP consumers. Overall, we demonstrate how extracellular fluxes quantitatively reflect intracellular ATP turnover and cellular bioenergetics. We provide a simple spreadsheet to calculate glycolytic and oxidative ATP production rates from raw extracellular acidification and respiration data. PMID:28270511

Multiscale approach to link red blood cell dynamics, shear viscosity, and ATP release.

PubMed

Forsyth, Alison M; Wan, Jiandi; Owrutsky, Philip D; Abkarian, Manouk; Stone, Howard A

2011-07-05

RBCs are known to release ATP, which acts as a signaling molecule to cause dilation of blood vessels. A reduction in the release of ATP from RBCs has been linked to diseases such as type II diabetes and cystic fibrosis. Furthermore, reduced deformation of RBCs has been correlated with myocardial infarction and coronary heart disease. Because ATP release has been linked to cell deformation, we undertook a multiscale approach to understand the links between single RBC dynamics, ATP release, and macroscopic viscosity all at physiological shear rates. Our experimental approach included microfluidics, ATP measurements using a bioluminescent reaction, and rheology. Using microfluidics technology with high-speed imaging, we visualize the deformation and dynamics of single cells, which are known to undergo motions such as tumbling, swinging, tanktreading, and deformation. We report that shear thinning is not due to cellular deformation as previously believed, but rather it is due to the tumbling-to-tanktreading transition. In addition, our results indicate that ATP release is constant at shear stresses below a threshold (3 Pa), whereas above the threshold ATP release is increased and accompanied by large cellular deformations. Finally, performing experiments with well-known inhibitors, we show that the Pannexin 1 hemichannel is the main avenue for ATP release both above and below the threshold, whereas, the cystic fibrosis transmembrane conductance regulator only contributes to deformation-dependent ATP release above the stress threshold.

Role of connexin 32 hemichannels in the release of ATP from peripheral nerves.

PubMed

Nualart-Marti, Anna; del Molino, Ezequiel Mas; Grandes, Xènia; Bahima, Laia; Martin-Satué, Mireia; Puchal, Rafel; Fasciani, Ilaria; González-Nieto, Daniel; Ziganshin, Bulat; Llobet, Artur; Barrio, Luis C; Solsona, Carles

2013-12-01

Extracellular purines elicit strong signals in the nervous system. Adenosine-5'-triphosphate (ATP) does not spontaneously cross the plasma membrane, and nervous cells secrete ATP by exocytosis or through plasma membrane proteins such as connexin hemichannels. Using a combination of imaging, luminescence and electrophysiological techniques, we explored the possibility that Connexin 32 (Cx32), expressed in Schwann cells (SCs) myelinating the peripheral nervous system could be an important source of ATP in peripheral nerves. We triggered the release of ATP in vivo from mice sciatic nerves by electrical stimulation and from cultured SCs by high extracellular potassium concentration-evoked depolarization. No ATP was detected in the extracellular media after treatment of the sciatic nerve with Octanol or Carbenoxolone, and ATP release was significantly inhibited after silencing Cx32 from SCs cultures. We investigated the permeability of Cx32 to ATP by expressing Cx32 hemichannels in Xenopus laevis oocytes. We found that ATP release is coupled to the inward tail current generated after the activation of Cx32 hemichannels by depolarization pulses, and it is sensitive to low extracellular calcium concentrations. Moreover, we found altered ATP release in mutated Cx32 hemichannels related to the X-linked form of Charcot-Marie-Tooth disease, suggesting that purinergic-mediated signaling in peripheral nerves could underlie the physiopathology of this neuropathy. Copyright © 2013 Wiley Periodicals, Inc.

Monitoring of the ADP/ATP Ratio by Induced Circularly Polarised Europium Luminescence.

PubMed

Shuvaev, Sergey; Fox, Mark A; Parker, David

2018-06-18

A series of three europium complexes bearing picolyl amine moieties was found to possess differing binding affinities towards Zn 2+ and three nucleotides: AMP, ADP, and ATP. A large increase in the total emission intensity was observed upon binding Zn 2+ , followed by signal amplification upon the addition of nucleotides. The resulting adducts possessed strong induced circularly polarised emission, with ADP and ATP signals of opposite sign. Model DFT geometries of the adducts suggest the Δ diastereoisomer is preferred for ATP and the Λ isomer for ADP/AMP. This change in sign allows the ADP/ATP (or AMP/ATP) ratio to be assessed by monitoring changes in the emission dissymmetry factor, g em . © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

ATP-induced current in isolated outer hair cells of guinea pig cochlea.

PubMed

Nakagawa, T; Akaike, N; Kimitsuki, T; Komune, S; Arima, T

1990-05-01

1. Electrical and pharmacologic properties of ATP-induced current in outer hair cells isolated from guinea pig cochlea were investigated in the whole-cell recording mode by the use of a conventional patch-clamp technique. 2. Under current-clamp conditions, rapid application of ATP depolarized the outer hair cells resulting in an increase in conductance. The ATP-induced response did not show any desensitization during a continuous application. 3. At a holding potential of -70 mV, the ATP-induced inward current increased in a sigmoidal fashion over the concentration range between 3 microM and 1 mM. The half-maximum concentration (EC50) was 12 microM and the Hill coefficient was 0.93. 4. The ATP-induced current had a reversal potential near 6 mV, which was close to the theoretical value (1 mV) calculated from the Goldman-Hodgkin-Katz equation for permeable intra- and extracellular cations. 5. In the current-voltage (I-V) relationship for the ATP response, a slight inward-going rectification was observed at more positive potentials than the reversal potential. 6. The substitution of extracellular Na+ by equimolar choline+ shifted the reversal potential of the ATP-induced current to more negative values. The substitution of Cs+ in the internal solution by N-methyl-D-glucamine+ (NMG+) shifted it in the positive direction. The reversal potential of ATP-induced current was also shifted to positive values with increasing extracellular Ca2+ concentration. A decrease of intracellular Cl- by gluconate- did not affect the reversal potential, thereby indicating that the ATP-induced current is carried through a large cation channel.(ABSTRACT TRUNCATED AT 250 WORDS)

Ionotropic P2X ATP Receptor Channels Mediate Purinergic Signaling in Mouse Odontoblasts

PubMed Central

Shiozaki, Yuta; Sato, Masaki; Kimura, Maki; Sato, Toru; Tazaki, Masakazu; Shibukawa, Yoshiyuki

2017-01-01

ATP modulates various functions in the dental pulp cells, such as intercellular communication and neurotransmission between odontoblasts and neurons, proliferation of dental pulp cells, and odontoblast differentiation. However, functional expression patterns and their biophysical properties of ionotropic ATP (P2X) receptors (P2X1–P2X7) in odontoblasts were still unclear. We examined these properties of P2X receptors in mouse odontoblasts by patch-clamp recordings. K+-ATP, nonselective P2X receptor agonist, induced inward currents in odontoblasts in a concentration-dependent manner. K+-ATP-induced currents were inhibited by P2X4 and P2X7 selective inhibitors (5-BDBD and KN62, respectively), while P2X1 and P2X3 inhibitors had no effects. P2X7 selective agonist (BzATP) induced inward currents dose-dependently. We could not observe P2X1, 2/3, 3 selective agonist (αβ-MeATP) induced currents. Amplitudes of K+-ATP-induced current were increased in solution without extracellular Ca2+, but decreased in Na+-free extracellular solution. In the absence of both of extracellular Na+ and Ca2+, K+-ATP-induced currents were completely abolished. K+-ATP-induced Na+ currents were inhibited by P2X7 inhibitor, while the Ca2+ currents were sensitive to P2X4 inhibitor. These results indicated that odontoblasts functionally expressed P2X4 and P2X7 receptors, which might play an important role in detecting extracellular ATP following local dental pulp injury. PMID:28163685

Ionotropic P2X ATP Receptor Channels Mediate Purinergic Signaling in Mouse Odontoblasts.

PubMed

Shiozaki, Yuta; Sato, Masaki; Kimura, Maki; Sato, Toru; Tazaki, Masakazu; Shibukawa, Yoshiyuki

2017-01-01

ATP modulates various functions in the dental pulp cells, such as intercellular communication and neurotransmission between odontoblasts and neurons, proliferation of dental pulp cells, and odontoblast differentiation. However, functional expression patterns and their biophysical properties of ionotropic ATP (P2X) receptors (P2X 1 -P2X 7 ) in odontoblasts were still unclear. We examined these properties of P2X receptors in mouse odontoblasts by patch-clamp recordings. K + -ATP, nonselective P2X receptor agonist, induced inward currents in odontoblasts in a concentration-dependent manner. K + -ATP-induced currents were inhibited by P2X 4 and P2X 7 selective inhibitors (5-BDBD and KN62, respectively), while P2X 1 and P2X 3 inhibitors had no effects. P2X 7 selective agonist (BzATP) induced inward currents dose-dependently. We could not observe P2X 1, 2/3, 3 selective agonist (αβ-MeATP) induced currents. Amplitudes of K + -ATP-induced current were increased in solution without extracellular Ca 2+ , but decreased in Na + -free extracellular solution. In the absence of both of extracellular Na + and Ca 2+ , K + -ATP-induced currents were completely abolished. K + -ATP-induced Na + currents were inhibited by P2X 7 inhibitor, while the Ca 2+ currents were sensitive to P2X 4 inhibitor. These results indicated that odontoblasts functionally expressed P2X 4 and P2X 7 receptors, which might play an important role in detecting extracellular ATP following local dental pulp injury.

The molecular motor F-ATP synthase is targeted by the tumoricidal protein HAMLET.

PubMed

Ho, James; Sielaff, Hendrik; Nadeem, Aftab; Svanborg, Catharina; Grüber, Gerhard

2015-05-22

HAMLET (human alpha-lactalbumin made lethal to tumor cells) interacts with multiple tumor cell compartments, affecting cell morphology, metabolism, proteasome function, chromatin structure and viability. This study investigated if these diverse effects of HAMLET might be caused, in part, by a direct effect on the ATP synthase and a resulting reduction in cellular ATP levels. A dose-dependent reduction in cellular ATP levels was detected in A549 lung carcinoma cells, and by confocal microscopy, co-localization of HAMLET with the nucleotide-binding subunits α (non-catalytic) and β (catalytic) of the energy converting F1F0 ATP synthase was detected. As shown by fluorescence correlation spectroscopy, HAMLET binds to the F1 domain of the F1F0 ATP synthase with a dissociation constant (KD) of 20.5μM. Increasing concentrations of the tumoricidal protein HAMLET added to the enzymatically active α3β3γ complex of the F-ATP synthase lowered its ATPase activity, demonstrating that HAMLET binding to the F-ATP synthase effects the catalysis of this molecular motor. Single-molecule analysis was applied to study HAMLET-α3β3γ complex interaction. Whereas the α3β3γ complex of the F-ATP synthase rotated in a counterclockwise direction with a mean rotational rate of 3.8±0.7s(-1), no rotation could be observed in the presence of bound HAMLET. Our findings suggest that direct effects of HAMLET on the F-ATP synthase may inhibit ATP-dependent cellular processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Purinergic and muscarinic modulation of ATP release from the urothelium and its paracrine actions

PubMed Central

Sui, Guiping; Fry, Chris H.; Montgomery, Bruce; Roberts, Max; Wu, Rui

2013-01-01

The urothelium is a newly recognized sensory structure that detects bladder fullness. Pivotal to this sensory role is the release of ATP from the urothelium. However, the routes for urothelial ATP release, its modulation by receptor-mediated pathways, and the autocrine/paracrine role of ATP are poorly understood, especially in native tissue. We examined the action of key neurotransmitters: purinergic and muscarinic agonists on ATP release and its paracrine effect. Guinea pig and human urothelial mucosa were mounted in a perfusion trough; superfusate ATP was measured using a luciferin-luciferase assay, and tissue contractions were recorded with a tension transducer. Intracellular Ca2+ was measured in isolated urothelial cells with fura-2. The P2Y agonist UTP but not the P2X agonist α,β-methylene-ATP generated ATP release. The muscarinic agonist carbachol and the M2-preferential agonist oxotremorine also generated ATP release, which was antagonized by the M2-specific agent methoctramine. Agonist-evoked ATP release was accompanied by mucosal contractions. Urothelial ATP release was differentially mediated by intracellular Ca2+ release, cAMP, exocytosis, or connexins. Urothelium-attached smooth muscle exhibited spontaneous contractions that were augmented by subthreshold concentrations of carbachol, which had little direct effect on smooth muscle. This activity was attenuated by desensitizing P2X receptors on smooth muscle. Urothelial ATP release was increased in aging bladders. Purinergic and muscarinic agents produced similar effects in human urothelial tissue. This is the first demonstration of specific modulation of urothelial ATP release in native tissue by purinergic and muscarinic neurotransmitters via distinct mechanisms. Released ATP produces paracrine effects on underlying tissues. This process is altered during aging and has relevance to human bladder pathologies. PMID:24285497

Purinergic and muscarinic modulation of ATP release from the urothelium and its paracrine actions.

PubMed

Sui, Guiping; Fry, Chris H; Montgomery, Bruce; Roberts, Max; Wu, Rui; Wu, Changhao

2014-02-01

The urothelium is a newly recognized sensory structure that detects bladder fullness. Pivotal to this sensory role is the release of ATP from the urothelium. However, the routes for urothelial ATP release, its modulation by receptor-mediated pathways, and the autocrine/paracrine role of ATP are poorly understood, especially in native tissue. We examined the action of key neurotransmitters: purinergic and muscarinic agonists on ATP release and its paracrine effect. Guinea pig and human urothelial mucosa were mounted in a perfusion trough; superfusate ATP was measured using a luciferin-luciferase assay, and tissue contractions were recorded with a tension transducer. Intracellular Ca²⁺ was measured in isolated urothelial cells with fura-2. The P2Y agonist UTP but not the P2X agonist α,β-methylene-ATP generated ATP release. The muscarinic agonist carbachol and the M₂-preferential agonist oxotremorine also generated ATP release, which was antagonized by the M₂-specific agent methoctramine. Agonist-evoked ATP release was accompanied by mucosal contractions. Urothelial ATP release was differentially mediated by intracellular Ca²⁺ release, cAMP, exocytosis, or connexins. Urothelium-attached smooth muscle exhibited spontaneous contractions that were augmented by subthreshold concentrations of carbachol, which had little direct effect on smooth muscle. This activity was attenuated by desensitizing P2X receptors on smooth muscle. Urothelial ATP release was increased in aging bladders. Purinergic and muscarinic agents produced similar effects in human urothelial tissue. This is the first demonstration of specific modulation of urothelial ATP release in native tissue by purinergic and muscarinic neurotransmitters via distinct mechanisms. Released ATP produces paracrine effects on underlying tissues. This process is altered during aging and has relevance to human bladder pathologies.

Inflight source noise of an advanced full-scale single-rotation propeller

NASA Technical Reports Server (NTRS)

Woodward, Richard P.; Loeffler, Irvin J.

1991-01-01

Flight tests to define the far field tone source at cruise conditions were completed on the full scale SR-7L advanced turboprop which was installed on the left wing of a Gulfstream II aircraft. This program, designated Propfan Test Assessment (PTA), involved aeroacoustic testing of the propeller over a range of test conditions. These measurements defined source levels for input into long distance propagation models to predict en route noise. Inflight data were taken for 7 test cases. The sideline directivities measured by the Learjet showed expected maximum levels near 105 degrees from the propeller upstream axis. However, azimuthal directivities based on the maximum observed sideline tone levels showed highest levels below the aircraft. An investigation of the effect of propeller tip speed showed that the tone level of reduction associated with reductions in propeller tip speed is more significant in the horizontal plane than below the aircraft.

Interactions of diamines with adenosine-5'-triphosphate (ATP) in the systems including copper(II) ions.

PubMed

Bregier-Jarzebowska, R; Gasowska, A; Hoffmann, S K; Lomozik, L

2016-09-01

Interactions were studied in the systems ATP/tn and ATP/Put (tn=1,3-diaminopropane, Put=putrescine) whereas the complexation reactions in ternary systems Cu(II)/ATP/tn and Cu(II)/ATP/Put. Results of the potentiometric and spectroscopic studies evidenced the formation of adducts of the type (ATP)H x (PA), where PA=diamine. The thermodynamic stability of the complexes and the mode of interactions were determined. On the basis of analysis of changes in the positions of NMR signals, in the pH range of (ATP)H 3 (Put) formation, the preferred centres of the interaction between ATP and Put are the endocyclic nitrogen atoms from the nucleotide. On the other hand, the shorter diamine tn in the entire pH range reacts with the phosphate groups from ATP. The positive centres of noncovalent interactions are the protonated NH x + groups from amines. In both complexes Cu(ATP)H 2 (tn) and Cu(ATP)H 3 (Put) formed in ternary systems at pH<6.5, the amines are in the outer sphere of coordination with the noncovalent interaction with anchoring Cu(ATP). Only the phosphate groups from the nucleotide take part in metalation. At higher pH in the range of Cu(ATP)(PA) complex formation, significant differences in the reactions of the two amines appear. The shorter one (tn) binds Cu(II) ions with two nitrogen atoms, while putrescine coordinates in the monofunctional mode, which is undoubtedly related to the differences in lengths of methylene chain. This explains the considerable differences in the stability of Cu(ATP)(tn) and Cu(ATP)(Put). In both complexes the nucleotide is coordinated through phosphate groups. As a result of noncovalent interactions ATP forms molecular complexes with 1,3-diaminopropane and 1,4-diaminobutane (putrescine). Significant differences in the mode of interactions between the two diamines were observed in ATP/diamine binary systems and in ternary systems Cu(II)/ATP/diamine, at high pH. Copyright © 2016 Elsevier Inc. All rights reserved.

Protecting muscle ATP: positive roles for peripheral defense mechanisms-introduction.

PubMed

Myburgh, Kathryn H

2004-01-01

Skeletal muscle has evolved an impressive array of mechanisms for peripherally mediated control of ATP homeostasis. Some of these mechanisms are intracellular, and others are extracellular and include influences on the cross-bridge cycle itself and substrate supply. This paper introduces three distinctly different topics that nevertheless all have ATP defense in common. The role of ADP in fatigue is controversial but has recently been more clearly delineated so that an effect on alleviating force declines during extreme fatigue is plausible. AMP plays its role by activating the protein-kinase, AMPK, which is a key sensor of cellular energy stress. AMPK has different isoforms, is not uniformly distributed in the cell, and its activation is carefully controlled. It has multiple effects including improvements in substrate supply for the metabolic pathways producing ATP and inhibition of anabolic processes to further spare ATP. Red blood cells have the capacity to sense hypoxia and to release vasodilators where there is a locally increased demand for blood supply. The papers in this series emphasize the important positive roles of metabolites and sensors of fatigue in the balance between ATP supply and demand.

Adenosine monophosphate as a mediator of ATP effects at P1 purinoceptors

PubMed Central

Ross, Fiona M; Brodie, Martin J; Stone, Trevor W

1998-01-01

When perfused with a medium containing no added magnesium and 4-aminopyridine (4AP) (50 μM) hippocampal slices generated epileptiform bursts of an interictal nature. We have shown in a previous study that adenosine 5′-triphosphate (ATP) depressed epileptiform activity and that this effect was blocked by the adenosine A1 receptor antagonist cyclopentyltheophylline but was not affected by adenosine deaminase. This implied that ATP might act indirectly at P1 receptors or at a xanthine-sensitive P2 receptor. The aim of the present study was to investigate further the action of ATP on epileptiform activity.ATP can be metabolized by ecto-nucleotidases to adenosine 5′-diphosphate (ADP), adenosine 5′-monophosphate (AMP) and adenosine, respectively. Each of these metabolites can activate receptors in its own right: P2 receptors for ADP and P1 receptors for AMP and adenosine.We now show that both AMP and ATP (50 μM) significantly decrease epileptiform discharge rate in a rapid and reversible manner. 5′Adenylic acid deaminase (AMP deaminase, AMPase) (0.2 u ml−1), when perfused alone did not significantly alter the discharge rate over the 10 min superfusion period used for drug application. When perfused concurrently with AMP (50 μM), AMP deaminase prevented the depressant effect of AMP on discharge rate.AMP deaminase, at a concentration of 0.2 u ml−1 which annulled the effect of AMP (50 μM), prevented the inhibitory activity of ATP (50 μM). A higher concentration of ATP (200 μM) depressed the frequency of spontaneous bursts to approximately 30% control and this response was also prevented by AMP deaminase.Superfusion of the slices with 5′-nucleotidase also prevented the inhibitory activity of ATP on epileptiform discharges.The results suggest that AMP mediates the inhibitory effects of ATP on epileptiform activity, a conclusion which can explain the earlier finding that cyclopentyltheophylline but not adenosine deaminase inhibited the

MRP transporters as membrane machinery in the bradykinin-inducible export of ATP.

PubMed

Zhao, Yumei; Migita, Keisuke; Sun, Jing; Katsuragi, Takeshi

2010-04-01

Adenosine triphosphate (ATP) plays the role of an autocrine/paracrine signal molecule in a variety of cells. So far, however, the membrane machinery in the export of intracellular ATP remains poorly understood. Activation of B2-receptor with bradykinin-induced massive release of ATP from cultured taenia coli smooth muscle cells. The evoked release of ATP was unaffected by gap junction hemichannel blockers, such as 18alpha-glycyrrhetinic acid and Gap 26. Furthermore, the cystic fibrosis transmembrane regulator (CFTR) coupled Cl(-) channel blockers, CFTR(inh)172, 5-nitro-2-(3-phenylpropylamino)-benzoic acid, Gd3(+) and glibenclamide, failed to suppress the export of ATP by bradykinin. On the other, the evoked release of ATP was greatly reduced by multidrug resistance protein (MRP) transporter inhibitors, MK-571, indomethacin, and benzbromarone. From western blotting analysis, blots of MRP 1 protein only, but not MRP 2 and MRP 3 protein, appeared at 190 kD. However, the MRP 1 protein expression was not enhanced after loading with 1 muM bradykinin for 5 min. Likewise, niflumic acid and fulfenamic acid, Ca2(+)-activated Cl(-) channel blockers, largely abated the evoked release of ATP. The possibility that the MRP transporter system couples with Ca2(+)-activated Cl(-) channel activities is discussed here. These findings suggest that MRP transporters, probably MRP 1, unlike CFTR-Cl(-) channels and gap junction hemichannels, may contribute as membrane machinery to the export of ATP induced by G-protein-coupled receptor stimulation.

Propulsion/ACEE

NASA Technical Reports Server (NTRS)

1981-01-01

The research objectives of the NASA aircraft energy efficiency program are summarized. Engine component improvements for turbofan engines, diagnostics, the development of advanced turboprop engines, and propeller noise analysis are discussed.

Twisting and subunit rotation in single FOF1-ATP synthase

PubMed Central

Sielaff, Hendrik; Börsch, Michael

2013-01-01

FOF1-ATP synthases are ubiquitous proton- or ion-powered membrane enzymes providing ATP for all kinds of cellular processes. The mechanochemistry of catalysis is driven by two rotary nanomotors coupled within the enzyme. Their different step sizes have been observed by single-molecule microscopy including videomicroscopy of fluctuating nanobeads attached to single enzymes and single-molecule Förster resonance energy transfer. Here we review recent developments of approaches to monitor the step size of subunit rotation and the transient elastic energy storage mechanism in single FOF1-ATP synthases. PMID:23267178

Interaction between ATP, metal ions, glycine, and several minerals

NASA Technical Reports Server (NTRS)

Rishpon, J.; Ohara, P. J.; Lawless, J. G.; Lahav, N.

1982-01-01

Interactions between ATP, glycine and montmorillonite and kaolinite clay minerals in the presence of various metal cations are investigated. The adsorption of adenine nucleotides on clays and Al(OH)3 was measured as a function of pH, and glycine condensation was followed in the presence of ATP, ZnCl2, MgCl2 and either kaolinite or montmorillonite. The amounts of ATP and ADP adsorbed are found to decrease with increasing Ph, and to be considerably enhanced in experiments with Mg(2+)- and Zn(2+)-montmorillonite with respect to Na(+)-montmorillonite. The effects of divalent cations are less marked in kaolinite. Results for Al(OH)3 show the importance of adsorption at clay platelet edges at high pH. The decomposition of ATP during drying at high temperature is observed to be inhibited by small amounts of clay, vacuum, or Mg(2+) or Zn(2+) ions, and to be accompanied by peptide formation in the presence of glycine. Results suggest the importance of Zn(2+) and Mg(2+) in chemical evolution.

Annoyance caused by aircraft en route noise

NASA Technical Reports Server (NTRS)

Mccurdy, David A.

1992-01-01

A laboratory experiment was conducted to quantify the annoyance response of people on the ground to enroute noise generated by aircraft at cruise conditions. The en route noises were ground level recordings of eight advanced turboprop aircraft flyovers and six conventional turbofan flyovers. The eight advanced turboprop enroute noises represented the NASA Propfan Test Assessment aircraft operating at different combinations of altitude, aircraft Mach number, and propeller tip speed. The conventional turbofan en route noises represented six different commercial airliners. The overall durations of the en route noises varied from approximately 40 to 160 sec. In the experiment, 32 subjects judged the annoyance of the en route noises as well as recordings of the takeoff and landing noises of each of 5 conventional turboprop and 5 conventional turbofan aircraft. Each of the noises was presented at three sound pressure levels to the subjects in an anechoic listening room. Analysis of the judgments found small differences in annoyance between three combinations of aircraft type and operation. Current tone and corrections did not significantly improve en route annoyance prediction. The optimum duration-correction magnitude for en route noise was approximately 1 dB per doubling of effective duration.

Annoyance caused by aircraft en route noise

NASA Astrophysics Data System (ADS)

McCurdy, David A.

1992-03-01

A laboratory experiment was conducted to quantify the annoyance response of people on the ground to enroute noise generated by aircraft at cruise conditions. The en route noises were ground level recordings of eight advanced turboprop aircraft flyovers and six conventional turbofan flyovers. The eight advanced turboprop enroute noises represented the NASA Propfan Test Assessment aircraft operating at different combinations of altitude, aircraft Mach number, and propeller tip speed. The conventional turbofan en route noises represented six different commercial airliners. The overall durations of the en route noises varied from approximately 40 to 160 sec. In the experiment, 32 subjects judged the annoyance of the en route noises as well as recordings of the takeoff and landing noises of each of 5 conventional turboprop and 5 conventional turbofan aircraft. Each of the noises was presented at three sound pressure levels to the subjects in an anechoic listening room. Analysis of the judgments found small differences in annoyance between three combinations of aircraft type and operation. Current tone and corrections did not significantly improve en route annoyance prediction. The optimum duration-correction magnitude for en route noise was approximately 1 dB per doubling of effective duration.

An ATP sensitive light addressable biosensor for extracellular monitoring of single taste receptor cell.

PubMed

Wu, Chunsheng; Du, Liping; Zou, Ling; Zhao, Luhang; Wang, Ping

2012-12-01

Adenosine triphosphate (ATP) is considered as the key neurotransmitter in taste buds for taste signal transmission and processing. Measurements of ATP secreted from single taste receptor cell (TRC) with high sensitivity and specificity are essential for investigating mechanisms underlying taste cell-to-cell communications. In this study, we presented an aptamer-based biosensor for the detection of ATP locally secreted from single TRC. ATP sensitive DNA aptamer was used as recognition element and its DNA competitor was served as signal transduction element that was covalently immobilized on the surface of light addressable potentiometric sensor (LAPS). Due to the light addressable capability of LAPS, local ATP secretion from single TRC can be detected by monitoring the working potential shifts of LAPS. The results show this biosensor can detect ATP with high sensitivity and specificity. It is demonstrated this biosensor can effectively detect the local ATP secretion from single TRC responding to tastant mixture. This biosensor could provide a promising new tool for the research of taste cell-to-cell communications as well as for the detection of local ATP secretion from other types of ATP secreting individual cells.

ATP sensing in living plant cells reveals tissue gradients and stress dynamics of energy physiology

PubMed Central

De Col, Valentina; Fuchs, Philippe; Nietzel, Thomas; Elsässer, Marlene; Voon, Chia Pao; Candeo, Alessia; Seeliger, Ingo; Fricker, Mark D; Grefen, Christopher; Møller, Ian Max; Bassi, Andrea; Lim, Boon Leong; Zancani, Marco; Meyer, Andreas J; Costa, Alex; Wagner, Stephan; Schwarzländer, Markus

2017-01-01

Growth and development of plants is ultimately driven by light energy captured through photosynthesis. ATP acts as universal cellular energy cofactor fuelling all life processes, including gene expression, metabolism, and transport. Despite a mechanistic understanding of ATP biochemistry, ATP dynamics in the living plant have been largely elusive. Here, we establish MgATP2- measurement in living plants using the fluorescent protein biosensor ATeam1.03-nD/nA. We generate Arabidopsis sensor lines and investigate the sensor in vitro under conditions appropriate for the plant cytosol. We establish an assay for ATP fluxes in isolated mitochondria, and demonstrate that the sensor responds rapidly and reliably to MgATP2- changes in planta. A MgATP2- map of the Arabidopsis seedling highlights different MgATP2- concentrations between tissues and within individual cell types, such as root hairs. Progression of hypoxia reveals substantial plasticity of ATP homeostasis in seedlings, demonstrating that ATP dynamics can be monitored in the living plant. DOI: http://dx.doi.org/10.7554/eLife.26770.001 PMID:28716182

Modulation of nucleotide sensitivity of ATP-sensitive potassium channels by phosphatidylinositol-4-phosphate 5-kinase.

PubMed

Shyng, S L; Barbieri, A; Gumusboga, A; Cukras, C; Pike, L; Davis, J N; Stahl, P D; Nichols, C G

2000-01-18

ATP-sensitive potassium channels (K(ATP) channels) regulate cell excitability in response to metabolic changes. K(ATP) channels are formed as a complex of a sulfonylurea receptor (SURx), a member of the ATP-binding cassette protein family, and an inward rectifier K(+) channel subunit (Kir6.x). Membrane phospholipids, in particular phosphatidylinositol (PI) 4,5-bisphosphate (PIP(2)), activate K(ATP) channels and antagonize ATP inhibition of K(ATP) channels when applied to inside-out membrane patches. To examine the physiological relevance of this regulatory mechanism, we manipulated membrane PIP(2) levels by expressing either the wild-type or an inactive form of PI-4-phosphate 5-kinase (PIP5K) in COSm6 cells and examined the ATP sensitivity of coexpressed K(ATP) channels. Channels from cells expressing the wild-type PIP5K have a 6-fold lower ATP sensitivity (K(1/2), the half maximal inhibitory concentration, approximately 60 microM) than the sensitivities from control cells (K(1/2) approximately 10 microM). An inactive form of the PIP5K had little effect on the K(1/2) of wild-type channels but increased the ATP-sensitivity of a mutant K(ATP) channel that has an intrinsically lower ATP sensitivity (from K(1/2) approximately 450 microM to K(1/2) approximately 100 microM), suggesting a decrease in membrane PIP(2) levels as a consequence of a dominant-negative effect of the inactive PIP5K. These results show that PIP5K activity, which regulates PIP(2) and PI-3,4,5-P(3) levels, is a significant determinant of the physiological nucleotide sensitivity of K(ATP) channels.

Defining the Role of ATP Hydrolysis in Mitotic Segregation of Bacterial Plasmids

PubMed Central

Ah-Seng, Yoan; Rech, Jérôme; Lane, David; Bouet, Jean-Yves

2013-01-01

Hydrolysis of ATP by partition ATPases, although considered a key step in the segregation mechanism that assures stable inheritance of plasmids, is intrinsically very weak. The cognate centromere-binding protein (CBP), together with DNA, stimulates the ATPase to hydrolyse ATP and to undertake the relocation that incites plasmid movement, apparently confirming the need for hydrolysis in partition. However, ATP-binding alone changes ATPase conformation and properties, making it difficult to rigorously distinguish the substrate and cofactor roles of ATP in vivo. We had shown that mutation of arginines R36 and R42 in the F plasmid CBP, SopB, reduces stimulation of SopA-catalyzed ATP hydrolysis without changing SopA-SopB affinity, suggesting the role of hydrolysis could be analyzed using SopA with normal conformational responses to ATP. Here, we report that strongly reducing SopB-mediated stimulation of ATP hydrolysis results in only slight destabilization of mini-F, although the instability, as well as an increase in mini-F clustering, is proportional to the ATPase deficit. Unexpectedly, the reduced stimulation also increased the frequency of SopA relocation over the nucleoid. The increase was due to drastic shortening of the period spent by SopA at nucleoid ends; average speed of migration per se was unchanged. Reduced ATP hydrolysis was also associated with pronounced deviations in positioning of mini-F, though time-averaged positions changed only modestly. Thus, by specifically targeting SopB-stimulated ATP hydrolysis our study reveals that even at levels of ATPase which reduce the efficiency of splitting clusters and the constancy of plasmid positioning, SopB still activates SopA mobility and plasmid positioning, and sustains near wild type levels of plasmid stability. PMID:24367270

Glucose recruits K(ATP) channels via non-insulin-containing dense-core granules.

PubMed

Yang, Shao-Nian; Wenna, Nancy Dekki; Yu, Jia; Yang, Guang; Qiu, Hua; Yu, Lina; Juntti-Berggren, Lisa; Köhler, Martin; Berggren, Per-Olof

2007-09-01

beta cells rely on adenosine triphosphate-sensitive potassium (K(ATP)) channels to initiate and end glucose-stimulated insulin secretion through changes in membrane potential. These channels may also act as a constituent of the exocytotic machinery to mediate insulin release independent of their electrical function. However, the molecular mechanisms whereby the beta cell plasma membrane maintains an appropriate number of K(ATP) channels are not known. We now show that glucose increases K(ATP) current amplitude by increasing the number of K(ATP) channels in the beta cell plasma membrane. The effect was blocked by inhibition of protein kinase A (PKA) as well as by depletion of extracellular or intracellular Ca(2+). Furthermore, glucose promoted recruitment of the potassium inward rectifier 6.2 to the plasma membrane, and intracellular K(ATP) channels localized in chromogranin-positive/insulin-negative dense-core granules. Our data suggest that glucose can recruit K(ATP) channels to the beta cell plasma membrane via non-insulin-containing dense-core granules in a Ca(2+)- and PKA-dependent manner.

Mitochondrial Genome Integrity Mutations Uncouple the Yeast Saccharomyces cerevisiae ATP Synthase*║

PubMed Central

Wang, Yamin; Singh, Usha; Mueller, David M.

2013-01-01

The mitochondrial ATP synthase is a molecular motor, which couples the flow of rotons with phosphorylation of ADP. Rotation of the central stalk within the core of ATP synthase effects conformational changes in the active sites driving the synthesis of ATP. Mitochondrial genome integrity (mgi) mutations have been previously identified in the α-, β-, and γ-subunits of ATP synthase in yeast Kluyveromyces lactis and trypanosome Trypanosoma brucei. These mutations reverse the lethality of the loss of mitochondrial DNA in petite negative strains. Introduction of the homologous mutations in Saccharomyces cerevisiae results in yeast strains that lose mitochondrial DNA at a high rate and accompanied decreases in the coupling of the ATP synthase. The structure of yeast F1-ATPase reveals that the mgi residues cluster around the γ-subunit and selectively around the collar region of F1. These results indicate that residues within the mgi complementation group are necessary for efficient coupling of ATP synthase, possibly acting as a support to fix the axis of rotation of the central stalk. PMID:17244612

Primitive ATP-activated P2X receptors: discovery, function and pharmacology

PubMed Central

Fountain, Samuel J.

2013-01-01

Adenosine 5-triphosphate (ATP) is omnipresent in biology. It is therefore no surprise that organisms have evolved multifaceted roles for ATP, exploiting its abundance and restriction of passive diffusion across biological membranes. A striking role is the emergence of ATP as a bona fide transmitter molecule, whereby the movement of ATP across membranes serves as a chemical message through a direct ligand-receptor interaction. P2X receptors are ligand-gated ion channels that mediate fast responses to the transmitter ATP in mammalian cells including central and sensory neurons, vascular smooth muscle, endothelium, and leukocytes. Molecular cloning of P2X receptors and our understanding of structure-function relationships has provided sequence information with which to query an exponentially expanding wealth of genome sequence information including protist, early animal and human pathogen genomes. P2X receptors have now been cloned and characterized from a number of simple organisms. Such work has led to surprising new cellular roles for the P2X receptors family and an unusual phylogeny, with organisms such as Drosophila and C. elegans notably lacking P2X receptors despite retaining ionotropic receptors for other common transmitters that are present in mammals. This review will summarize current work on the evolutionary biology of P2X receptors and ATP as a signaling molecule, discuss what can be drawn from such studies when considering the action of ATP in higher animals and plants, and outline how simple organisms may be exploited experimentally to inform P2X receptor function in a wider context. PMID:24367292

Tomatidine Is a Lead Antibiotic Molecule That Targets Staphylococcus aureus ATP Synthase Subunit C.

PubMed

Lamontagne Boulet, Maxime; Isabelle, Charles; Guay, Isabelle; Brouillette, Eric; Langlois, Jean-Philippe; Jacques, Pierre-Étienne; Rodrigue, Sébastien; Brzezinski, Ryszard; Beauregard, Pascale B; Bouarab, Kamal; Boyapelly, Kumaraswamy; Boudreault, Pierre-Luc; Marsault, Éric; Malouin, François

2018-06-01

Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of deadly hospital-acquired infections. The discovery of anti- Staphylococcus antibiotics and new classes of drugs not susceptible to the mechanisms of resistance shared among bacteria is imperative. We recently showed that tomatidine (TO), a steroidal alkaloid from solanaceous plants, possesses potent antibacterial activity against S. aureus small-colony variants (SCVs), the notoriously persistent form of this bacterium that has been associated with recurrence of infections. Here, using genomic analysis of in vitro -generated TO-resistant S. aureus strains to identify mutations in genes involved in resistance, we identified the bacterial ATP synthase as the cellular target. Sequence alignments were performed to highlight the modified sequences, and the structural consequences of the mutations were evaluated in structural models. Overexpression of the atpE gene in S. aureus SCVs or introducing the mutation found in the atpE gene of one of the high-level TO-resistant S. aureus mutants into the Bacillus subtilis atpE gene provided resistance to TO and further validated the identity of the cellular target. FC04-100, a TO derivative which also possesses activity against non-SCV strains, prevents high-level resistance development in prototypic strains and limits the level of resistance observed in SCVs. An ATP synthesis assay allowed the observation of a correlation between antibiotic potency and ATP synthase inhibition. The selectivity index (inhibition of ATP production by mitochondria versus that of bacterial ATP synthase) is estimated to be >10 5 -fold for FC04-100. Copyright © 2018 American Society for Microbiology.

How the nucleus and mitochondria communicate in energy production during stress: nuclear MtATP6, an early-stress responsive gene, regulates the mitochondrial F₁F₀-ATP synthase complex.

PubMed

Moghadam, Ali Asghar; Ebrahimie, Eemaeil; Taghavi, Seyed Mohsen; Niazi, Ali; Babgohari, Mahbobeh Zamani; Deihimi, Tahereh; Djavaheri, Mohammad; Ramezani, Amin

2013-07-01

A small number of stress-responsive genes, such as those of the mitochondrial F1F0-ATP synthase complex, are encoded by both the nucleus and mitochondria. The regulatory mechanism of these joint products is mysterious. The expression of 6-kDa subunit (MtATP6), a relatively uncharacterized nucleus-encoded subunit of F0 part, was measured during salinity stress in salt-tolerant and salt-sensitive cultivated wheat genotypes, as well as in the wild wheat genotypes, Triticum and Aegilops using qRT-PCR. The MtATP6 expression was suddenly induced 3 h after NaCl treatment in all genotypes, indicating an early inducible stress-responsive behavior. Promoter analysis showed that the MtATP6 promoter includes cis-acting elements such as ABRE, MYC, MYB, GTLs, and W-boxes, suggesting a role for this gene in abscisic acid-mediated signaling, energy metabolism, and stress response. It seems that 6-kDa subunit, as an early response gene and nuclear regulatory factor, translocates to mitochondria and completes the F1F0-ATP synthase complex to enhance ATP production and maintain ion homeostasis under stress conditions. These communications between nucleus and mitochondria are required for inducing mitochondrial responses to stress pathways. Dual targeting of 6-kDa subunit may comprise as a mean of inter-organelle communication and save energy for the cell. Interestingly, MtATP6 showed higher and longer expression in the salt-tolerant wheat and the wild genotypes compared to the salt-sensitive genotype. Apparently, salt-sensitive genotypes have lower ATP production efficiency and weaker energy management than wild genotypes; a stress tolerance mechanism that has not been transferred to cultivated genotypes.

Diverse Functional Properties of Wilson Disease ATP7B Variants

PubMed Central

Huster, Dominik; Kühne, Angelika; Bhattacharjee, Ashima; Raines, Lily; Jantsch, Vanessa; Noe, Johannes; Schirrmeister, Wiebke; Sommerer, Ines; Sabri, Osama; Berr, Frieder; Mössner, Joachim; Stieger, Bruno; Caca, Karel; Lutsenko, Svetlana

2012-01-01

BACKGROUND & AIMS Wilson disease is a severe disorder of copper metabolism caused by mutations in ATP7B, which encodes a copper-transporting adenosine triphosphatase. The disease presents with a variable phenotype that complicates the diagnostic process and treatment. Little is known about the mechanisms that contribute to the different phenotypes of the disease. METHODS We analyzed 28 variants of ATP7B from patients with Wilson disease that affected different functional domains; the gene products were expressed using the baculovirus expression system in Sf9 cells. Protein function was analyzed by measuring catalytic activity and copper (64Cu) transport into vesicles. We studied intracellular localization of variants of ATP7B that had measurable transport activities and were tagged with green fluorescent protein in mammalian cells using confocal laser scanning microscopy. RESULTS Properties of ATP7B variants with pathogenic amino-acid substitution varied greatly even if substitutions were in the same functional domain. Some variants had complete loss of catalytic and transport activity, whereas others lost transport activity but retained phosphor-intermediate formation or had partial losses of activity. In mammalian cells, transport-competent variants differed in stability and subcellular localization. CONCLUSIONS Variants in ATP7B associated with Wilson disease disrupt the protein’s transport activity, result in its mislocalization, and reduce its stability. Single assays are insufficient to accurately predict the effects of ATP7B variants the function of its product and development of Wilson disease. These findings will contribute to our understanding of genotype–phenotype correlation and mechanisms of disease pathogenesis. PMID:22240481

Mutations in ATP6V1B1 and ATP6V0A4 genes cause recessive distal renal tubular acidosis in Mexican families.

PubMed

Escobar, Laura I; Simian, Christopher; Treard, Cyrielle; Hayek, Donia; Salvador, Carolina; Guerra, Norma; Matos, Mario; Medeiros, Mara; Enciso, Sandra; Camargo, María Dolores; Vargas-Poussou, Rosa

2016-05-01

Autosomal recessive distal renal tubular acidosis (dRTA) is a rare disease characterized by a hyperchloremic metabolic acidosis with normal anion gap, hypokalemia, hypercalciuria, hypocitraturia, nephrocalcinosis, and conserved glomerular filtration rate. In some cases, neurosensorial deafness is associated. dRTA is developed during the first months of life and the main manifestations are failure to thrive, vomiting, dehydration, and anorexia. Nine unrelated families were studied: seven children, a teenager, and an adult with dRTA. Hearing was preserved in four children. Coding regions of the genes responsible for recessive dRTA were analysed by Sanger sequencing. Molecular defects were found in the genes ATP6V1B1 and ATP6V0A4. We identified three homozygous variants in ATP6V1B: a frameshift mutation (p.Ile386Hisfs*56), a nucleotide substitution in exon 10 (p.Pro346Arg), and a new splicing mutation in intron 5. Three patients were homozygous for one novel (p.Arg743Trp) and one known (p.Asp411Tyr) missense mutations in the ATP6V0A4 gene. Three patients were compound heterozygous: one proband displayed two novel mutations, the frameshift mutation p.Val52Metfs*25, and a large deletion of exons 18-21; two probands showed the missense mutation p.Asp411Tyr and as a second mutation, p.Arg194Ter and c.1691+2dup, respectively. ATP6V0A4 and ATP6V1B1 genes were involved in recessive dRTA of Mexican families. All ATP6V1B1 mutations detected were homozygous and all patients developed sensorineural hearing loss (SNHL) early in infancy. ATP6V0A4 mutations were found in one infant and three children without SNHL, and in one teenager and one adult with SNHL confirming the phenotypic variability in this trait. The mutation p.Asp411Tyr detected in four Mexican families was due to a founder effect. Screening of these mutations could provide a rapid and valuable tool for diagnosis of dRTA in this population.

Online damage inspection of optics for ATP system

NASA Astrophysics Data System (ADS)

Chen, Jing; Jiang, Yu; Mao, Yao; Gan, Xun; Liu, Qiong

2016-09-01

In the Electro-Optical acquisition-tracking-pointing system (ATP), the optical components will be damaged with the several influencing factors. In this situation, the rate will increase sharply when the arrival of damage to some extent. As the complex processing techniques and long processing cycle of optical components, the damage will cause the great increase of the system development cost and cycle. Therefore, it is significant to detect the laser-induced damage in the ATP system. At present, the major research on the on-line damage detection technology of optical components is for the large optical system in the international. The relevant detection systems have complicated structures and many of components, and require enough installation space reserved, which do not apply for ATP system. To solve the problem mentioned before, This paper use a method based on machine vision to detect the damage on-line for the present ATP system. To start with, CCD and PC are used for image acquisition. Secondly, smoothing filters are used to restrain false damage points produced by noise. Then, with the shape feature included in the damage image, the OTSU Method which can define the best segmentation threshold automatically is used to achieve the goal to locate the damage regions. At last, we can supply some opinions for the lifetime of the optical components by analyzing the damage data, such as damage area, damage position. The method has the characteristics of few-detectors and simple-structures which can be installed without any changes of the original light path. With the method, experimental results show that it is stable and effective to achieve the goal of detecting the damage of optical components on-line in the ATP system.

Subtype-specific control of P2X receptor channel signaling by ATP and Mg2+.

PubMed

Li, Mufeng; Silberberg, Shai D; Swartz, Kenton J

2013-09-03

The identity and forms of activating ligands for ion channels are fundamental to their physiological roles in rapid electrical signaling. P2X receptor channels are ATP-activated cation channels that serve important roles in sensory signaling and inflammation, yet the active forms of the nucleotide are unknown. In physiological solutions, ATP is ionized and primarily found in complex with Mg(2+). Here we investigated the active forms of ATP and found that the action of MgATP(2-) and ATP(4-) differs between subtypes of P2X receptors. The slowly desensitizing P2X2 receptor can be activated by free ATP, but MgATP(2-) promotes opening with very low efficacy. In contrast, both free ATP and MgATP(2-) robustly open the rapidly desensitizing P2X3 subtype. A further distinction between these two subtypes is the ability of Mg(2+) to regulate P2X3 through a distinct allosteric mechanism. Importantly, heteromeric P2X2/3 channels present in sensory neurons exhibit a hybrid phenotype, characterized by robust activation by MgATP(2-) and weak regulation by Mg(2+). These results reveal the existence of two classes of homomeric P2X receptors with differential sensitivity to MgATP(2-) and regulation by Mg(2+), and demonstrate that both restraining mechanisms can be disengaged in heteromeric channels to form fast and sensitive ATP signaling pathways in sensory neurons.

Subtype-specific control of P2X receptor channel signaling by ATP and Mg2+

PubMed Central

Li, Mufeng; Silberberg, Shai D.; Swartz, Kenton J.

2013-01-01

The identity and forms of activating ligands for ion channels are fundamental to their physiological roles in rapid electrical signaling. P2X receptor channels are ATP-activated cation channels that serve important roles in sensory signaling and inflammation, yet the active forms of the nucleotide are unknown. In physiological solutions, ATP is ionized and primarily found in complex with Mg2+. Here we investigated the active forms of ATP and found that the action of MgATP2− and ATP4− differs between subtypes of P2X receptors. The slowly desensitizing P2X2 receptor can be activated by free ATP, but MgATP2− promotes opening with very low efficacy. In contrast, both free ATP and MgATP2− robustly open the rapidly desensitizing P2X3 subtype. A further distinction between these two subtypes is the ability of Mg2+ to regulate P2X3 through a distinct allosteric mechanism. Importantly, heteromeric P2X2/3 channels present in sensory neurons exhibit a hybrid phenotype, characterized by robust activation by MgATP2− and weak regulation by Mg2+. These results reveal the existence of two classes of homomeric P2X receptors with differential sensitivity to MgATP2− and regulation by Mg2+, and demonstrate that both restraining mechanisms can be disengaged in heteromeric channels to form fast and sensitive ATP signaling pathways in sensory neurons. PMID:23959888

The Mitochondrial Permeability Transition Pore: Channel Formation by F-ATP Synthase, Integration in Signal Transduction, and Role in Pathophysiology

PubMed Central

Bernardi, Paolo; Rasola, Andrea; Forte, Michael; Lippe, Giovanna

2015-01-01

The mitochondrial permeability transition (PT) is a permeability increase of the inner mitochondrial membrane mediated by a channel, the permeability transition pore (PTP). After a brief historical introduction, we cover the key regulatory features of the PTP and provide a critical assessment of putative protein components that have been tested by genetic analysis. The discovery that under conditions of oxidative stress the F-ATP synthases of mammals, yeast, and Drosophila can be turned into Ca2+-dependent channels, whose electrophysiological properties match those of the corresponding PTPs, opens new perspectives to the field. We discuss structural and functional features of F-ATP synthases that may provide clues to its transition from an energy-conserving into an energy-dissipating device as well as recent advances on signal transduction to the PTP and on its role in cellular pathophysiology. PMID:26269524

Molecular structure of human KATP in complex with ATP and ADP

PubMed Central

Lee, Kenneth Pak Kin

2017-01-01

In many excitable cells, KATP channels respond to intracellular adenosine nucleotides: ATP inhibits while ADP activates. We present two structures of the human pancreatic KATP channel, containing the ABC transporter SUR1 and the inward-rectifier K+ channel Kir6.2, in the presence of Mg2+ and nucleotides. These structures, referred to as quatrefoil and propeller forms, were determined by single-particle cryo-EM at 3.9 Å and 5.6 Å, respectively. In both forms, ATP occupies the inhibitory site in Kir6.2. The nucleotide-binding domains of SUR1 are dimerized with Mg2+-ATP in the degenerate site and Mg2+-ADP in the consensus site. A lasso extension forms an interface between SUR1 and Kir6.2 adjacent to the ATP site in the propeller form and is disrupted in the quatrefoil form. These structures support the role of SUR1 as an ADP sensor and highlight the lasso extension as a key regulatory element in ADP’s ability to override ATP inhibition. PMID:29286281

Visualization and Measurement of ATP Levels in Living Cells Replicating Hepatitis C Virus Genome RNA

PubMed Central

Ando, Tomomi; Imamura, Hiromi; Suzuki, Ryosuke; Aizaki, Hideki; Watanabe, Toshiki; Wakita, Takaji; Suzuki, Tetsuro

2012-01-01

Adenosine 5′-triphosphate (ATP) is the primary energy currency of all living organisms and participates in a variety of cellular processes. Although ATP requirements during viral lifecycles have been examined in a number of studies, a method by which ATP production can be monitored in real-time, and by which ATP can be quantified in individual cells and subcellular compartments, is lacking, thereby hindering studies aimed at elucidating the precise mechanisms by which viral replication energized by ATP is controlled. In this study, we investigated the fluctuation and distribution of ATP in cells during RNA replication of the hepatitis C virus (HCV), a member of the Flaviviridae family. We demonstrated that cells involved in viral RNA replication actively consumed ATP, thereby reducing cytoplasmic ATP levels. Subsequently, a method to measure ATP levels at putative subcellular sites of HCV RNA replication in living cells was developed by introducing a recently-established Förster resonance energy transfer (FRET)-based ATP indicator, called ATeam, into the NS5A coding region of the HCV replicon. Using this method, we were able to observe the formation of ATP-enriched dot-like structures, which co-localize with non-structural viral proteins, within the cytoplasm of HCV-replicating cells but not in non-replicating cells. The obtained FRET signals allowed us to estimate ATP concentrations within HCV replicating cells as ∼5 mM at possible replicating sites and ∼1 mM at peripheral sites that did not appear to be involved in HCV replication. In contrast, cytoplasmic ATP levels in non-replicating Huh-7 cells were estimated as ∼2 mM. To our knowledge, this is the first study to demonstrate changes in ATP concentration within cells during replication of the HCV genome and increased ATP levels at distinct sites within replicating cells. ATeam may be a powerful tool for the study of energy metabolism during replication of the viral genome. PMID:22396648

Spike-independent release of ATP from Xenopus spinal neurons evoked by activation of glutamate receptors

PubMed Central

Brown, Paul; Dale, Nicholas

2002-01-01

As the release of ATP from neurons has only been directly studied in a few cases, we have used patch sniffing to examine ATP release from Xenopus spinal neurons. ATP release was detected following intracellular current injection to evoke spikes. However, spiking was not essential as both glutamate and NMDA could evoke release of ATP in the presence of TTX. Neither acetylcholine nor high K+ was effective at inducing ATP release in the presence of TTX. Although Cd2+ blocked glutamate-evoked release of ATP suggesting a dependence on Ca2+ entry, neither ω-conotoxin-GVIA nor nifedipine prevented ATP release. N-type and L-type channels are thus not essential for glutamate-evoked ATP release. That glutamate receptors can elicit release in the absence of spiking suggests a close physical relationship between these receptors, the Ca2+ channels and release sites. As the dependence of ATP release on the influx of Ca2+ through Ca2+ channel subtypes differs from that of synaptic transmitter release, ATP may be released from sites that are distinct from those of the principal transmitter. In addition to its role as a fast transmitter, ATP may thus be released as a consequence of the activation of excitatory glutamatergic synapses and act to signal information about activity patterns in the nervous system. PMID:11986374

Controlled rotation of the F1-ATPase reveals differential and continuous binding changes for ATP synthesis

PubMed Central

Adachi, Kengo; Oiwa, Kazuhiro; Yoshida, Masasuke; Nishizaka, Takayuki; Kinosita, Kazuhiko

2012-01-01

F1-ATPase is an ATP-driven rotary molecular motor that synthesizes ATP when rotated in reverse. To elucidate the mechanism of ATP synthesis, we imaged binding and release of fluorescently labelled ADP and ATP while rotating the motor in either direction by magnets. Here we report the binding and release rates for each of the three catalytic sites for 360° of the rotary angle. We show that the rates do not significantly depend on the rotary direction, indicating ATP synthesis by direct reversal of the hydrolysis-driven rotation. ADP and ATP are discriminated in angle-dependent binding, but not in release. Phosphate blocks ATP binding at angles where ADP binding is essential for ATP synthesis. In synthesis rotation, the affinity for ADP increases by >104, followed by a shift to high ATP affinity, and finally the affinity for ATP decreases by >104. All these angular changes are gradual, implicating tight coupling between the rotor angle and site affinities. PMID:22929779

Modeling regulation of cardiac KATP and L-type Ca2+ currents by ATP, ADP, and Mg2+.

PubMed

Michailova, Anushka; Saucerman, Jeffrey; Belik, Mary Ellen; McCulloch, Andrew D

2005-03-01

Changes in cytosolic free Mg(2+) and adenosine nucleotide phosphates affect cardiac excitability and contractility. To investigate how modulation by Mg(2+), ATP, and ADP of K(ATP) and L-type Ca(2+) channels influences excitation-contraction coupling, we incorporated equations for intracellular ATP and MgADP regulation of the K(ATP) current and MgATP regulation of the L-type Ca(2+) current in an ionic-metabolic model of the canine ventricular myocyte. The new model: 1), quantitatively reproduces a dose-response relationship for the effects of changes in ATP on K(ATP) current, 2), simulates effects of ADP in modulating ATP sensitivity of K(ATP) channel, 3), predicts activation of Ca(2+) current during rapid increase in MgATP, and 4), demonstrates that decreased ATP/ADP ratio with normal total Mg(2+) or increased free Mg(2+) with normal ATP and ADP activate K(ATP) current, shorten action potential, and alter ionic currents and intracellular Ca(2+) signals. The model predictions are in agreement with experimental data measured under normal and a variety of pathological conditions.