Chemoelectrical Energy Conversion of Adenosine Triphosphate using ATPases

Abstract

Biological ion transport processes in proteins have inspired the development of bio-sensors, actuators, photoelectric, and chemoelectric energy conversion devices. These bio-inspired devices use ion transport through a protein energized by biochemical reactions in the protein’s sub-units to perform their engineering function. In an effort to advance the use of biological processes in synthetic systems, a chemoelectrical energy conversion device is demonstrated in this article that uses hydrolysis of adenosine triphosphate (ATP) in ATPase enzyme to generate electrical power. The ATPase enzyme in the device is reconstituted in a bilayer lipid membrane (BLM) and supported on a porous substrate. ATP is dissolved in pH7 buffer and added to one of the chambers in this bicameral device. The transmembrane gradient established by proton transport, resulting from hydrolysis of ATP in the enzyme, is converted into electron flow in an external circuit via silver-silver chloride electrodes placed in the buffer solution on both the sides of the membrane. The chemoelectrical energy conversion of ATP is demonstrated in this article using electrical impedance spectroscopy and load characterization experiments on BLMs supported in the pore(s) of a 25% porous polycarbonate membrane and single-pore silicon nitride chip. In electrical impedance spectroscopy, the change in conductance states of the membrane quantified by specific resistance is used to demonstrate protein activity. The mean electrical impedance of the BLM with ATPase supported on a single pore silicon nitride chip drops from 7 kΩ cm² to 250 Ω cm² on adding ATP to one side of the membrane. This change in ionic conductance of the BLM with ATPase in the presence of ATP demonstrates protein activity in the membrane. Impedance analysis of the membranes (BLM, BLM with ATPase) supported in multi-pore polycarbonate substrate demonstrates similar trend confirming ion transport and energy conversion in the membrane. In load characterization experiments, a resistive load of known magnitude is connected to the device and voltage across the membrane and current through the circuit are measured. The current−voltage characteristics of the device resembles a constant current power source and the slope of the response represents the internal resistance of the device. The current through the membrane supported on the single pore substrate is below the range of our data acquisition equipment and hence the device with the porous polycarbonate membrane is used in load characterization experiments. This polycarbonate membrane-based device has an open-circuit voltage of 87.5(±7.5) mV with a specific power output of 1.85 μW/cm² and an internal resistance of 30.5k(±8.5k)Ω. The theoretical maximum specific power available from the membrane supported on the multi-pore polycarbonate membrane and the single pore silicon-nitride substrate are estimated from the current−voltage response to be 7.65 and 18 μW/cm².

Keywords

membrane transport energy harvesting ATPase chemoelectrical energy conversion adenosine triphosphate.

Get full access to this article

View all access options for this article.

References

Akle, B. , Bennett, M. and Leo, D.J. 2006. ‘‘Development of a Novel Electrochemically Active Membrane and ‘Smart’ Material Based Vibration Sensor/Damper ,’’ Sensors and Actuators A: Physical, 126:173-181.

Arata, Y. , Nishi, T. , Kawasaki-Nishi, S. , Shao, E. , Wilkens, S. and Forgac, M. 2002. ‘‘Structure, Subunit Function and Regulation of the Coated Vesicle and Yeast Vacuolar (H+)-Atpases’’ , Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1555:71-74.

Bennett, M. and Leo, D. 2005. ‘‘Morphological and Electromechanical Characterization of Ionic Liquid/Nafion Polymer Composites,’’ In: Proceedings of the SPIE.

Bayley, H. , Radhakrishnan, R. and Huang, K. 1981. ‘‘Light-driven Proton Translocation by Bacteriorhodopsin Reconstituted with the Phenyl Analog of Retinal, ’’ Journal of Biological Chemistry, 256:3797-3801.

Buch-Pedersen, M.J. and Palmgren, M.G. 2003. ‘‘Mechanism of Proton Transport by Plant Plasma Membrane Proton ATPases,’’ Journal of Plant Research , 116:507-515.

Briskin, D.P. and Reynolds-Niesman, I. 1991. ‘‘Determination of H+/ ATP Stoichiometry for the Plasma Membrane H-ATPase from Red Beet (Beta vulgaris L.) Storage Tissue,’’ Plant Physiology, 95:242-250.

Briskin, D.P. 2005. ‘‘Intermediate Reaction States of the Red Beet Plasma Membrane ATPase,’’ Archives of Biochemistry and Biophysics, 248:106-115.

Beyenbach, K.W. and Wieczorek, H. 2006. ‘‘Review: The V-type H+ ATPase: Molecular Structure and Function, Physiological Roles and Regulation, ’’ The Journal of Experimental Biology, 209:577-589.

Carpaneto, A. , Geiger, D. , Bamberg, E. , Sauer, N. , Fromm, J. and Hedrich, R. 2005. ‘‘Phloem-localized, Proton-coupled Sucrose Carrier ZmSUT1 Mediates Sucrose Efflux under the Control of the Sucrose Gradient and the Proton Motive Force,’’ Journal of Biological Chemistry, 280:21437-21443.

10.

Cheng, Y. , Ogier, S.D. , Bushby, R.J. and Evans, S.D. 2000. ‘‘Discrete Membrane Arrays,’’ Reviews in Molecular Biotechnology, 74:159-174.

11.

Castellana, E.T. and Cremer, P.S. 2006. ‘‘Solid Supported Lipid Bilayers: From Biophysical Studies To Sensor Design,’’ Surface Science Reports , 61(10):429-444.

12.

Craighead, H. 2006. ‘‘Future Lab-on-a-chip Technologies for Interrogating Individual Molecules,’’ Nature , 442:387-393.

13.

Chandramohandas, R. 2006. ‘‘Rapid Purification Of Human Lysosomal Membranes, Characterisation of the Detergent Resistant Microdomains, and Purification and Reconstitution of the Vacuolar Proton Pump (V-ATPase),’’ PhD thesis.

14.

Dittrich, P. and Manz, A. 2006. ‘‘Lab-on-a-chip: Microuidics in Drug Discovery ,’’ Nature Reviews Drug Discovery, 5:210-218.

15.

Druzhko, A. , Vanderah, D. and Robertson, B. 1996. ‘‘An Azulenic Bacteriorhodopsin Analog has Photoinduced Activity,’’ Photochemistry and Photobiology , 64:867-869.

16.

Drachev, L.A. , Jasaitis, A.A. , Kaulen, A.D. , Kondrashin, A.A. , Liberman, E.A. , Nemecek, I.B. , Ostroumov, S.A. , Semenov, A.Y. and Skulachev, V.P. 1974. ‘‘Direct Measurement of Electric Current Generation by Cytochrome Oxidase, H+-ATPase and Bacteriorhodopsin,’’ Nature, 249: 321-324.

17.

Forgac, M. 1998. ‘‘Minireview - Structure, Function and Regulation of the Vacuolar H+-ATPases,’’ FEBS Letters, 440:258-263.

18.

Gu, L. , Wang, L. , Xun, J. and Ottova-Leitmannova, A. 1996. ‘‘A New Method for the Determination of Electrical Properties of Supported Bilayer Lipid Membranes by Cyclic Voltammetry , ’’ Bioelectrochemistry and Bioenergetics, 39:275-283.

19.

Ho, D. , Chu, B. and Montemagno, H.L.C.D. 2004. ‘‘Protein-driven Energy Transduction across Polymeric Biomembranes,’’ Nanotechnology , 15:1084-1094.

20.

Hess, H. , Bachand, G. and Vogel, V. 2004. ‘‘Powering Nanodevices with Biomolecular Motors ,’’ Chemistry-A European Journal, 10:2110-2116.

21.

Hess, H. 2006. ‘‘Self-assembly Driven by Molecular Motors ,’’ Soft Matter, 2:669-677.

22.

Hiraki, K. , Hamanaka, T. , Zheng, X. and Shinada, T. 2002. ‘‘Bacteriorhodopsin Analog Regenerated with 13-Desmethyl-13-Iodoretinal,’’ Biophysical Journal , 83:3460-3469.

23.

Hirata, H. , Ohno, K. , Sone, N. , Kagawa, Y. and Hamamoto, T. 1986. ‘‘Direct Measurement of the Electrogenicity of the H+-ATPase from Thermophilic Bacterium PS3 Reconstituted in Planar Phospholipid Bilayers,’’ The Journal of Biological Chemistry , 261:9839-9843.

24.

Hopkinson, D. 2007. ‘‘Measurements and Modeling of the Failure Pressure of Bilayer Lipid Membranes,’’ Available at: www.scholar.lib.vt.edu.

25.

Lanyi, J.K. 1993. ‘‘Pathways of Proton Transfer in the Light-driven Pump Bacteriorhodopsin,’’ Cellular and Molecular Life Sciences (CMLS), 49:514-517.

26.

Luecke, H. , Richter, H.-T. and Lanyi, J.K. 1998. ‘‘Proton Transfer Pathways in Bacteriorhodopsin at 2.3 Angstrom Resolution,’’ Science, 280:1934-1937.

27.

Mettler, I. and Leonard, R. 1979. ‘‘Ion Transport in Isolated Protoplasts from Tobacco Suspension Cells I. General,’’ Plant Physiology , 63:183-190.

28.

Medoro, G. , Manaresi, N. , Leonardi, A. and Altomare, L. 2003. ‘‘A Lab-on-a-chip for Cell Detection and Manipulation ,’’ IEEE Sensors Journal, 3:317-325.

29.

Nelson, N. and Taiz, L. 1989. ‘‘The Evolution of H+-ATPases,’’ Trends in Biochemical Sciences, 14:113-116.

30.

Nemat-Nasser, S. 2002. ‘‘Micromechanics of Actuation of Ionic Polymer-metal Composites,’’ Journal of Applied Physics, 92:2899-2915.

31.

Puri, N. , Lai-Zhang, J. , Meier, S. and Mueller, D.M. 2005. ‘‘Expression of Bovine F1-ATPase with Functional Complementation in Yeast Saccharomyces cerevisiae, ’’ Journal of Biological Chemistry, 280:22418-22424.

32.

Pancrazio, J.J. , Whelan, J.P. , Borkholder, D.A. , Ma, W. and Stenger, D.A. 1999. ‘‘Development and Application of Cell-based Biosensors,’’ Annals of Biomedical Engineering , 27:697-711.

33.

Ratajczak, R. 2000. ‘‘Review: Structure, Function and Regulation of the Plant Vacuolar H-translocating ATPases,’’ Biochimica Biophysica Acta, 1465:17-36.

34.

Sundaresan, V.B. and Leo, D.J. 2008. ‘‘Modeling and Characterization of a Chemomechanical Actuator using Protein Transporters,’’ Sensors and Actuators B: Chemical, 131:384-393.

35.

Shahinpoor, M. , Bar-Cohen, Y. , Simpson, J.O. and Smith, J. 1998. ‘‘Ionic Polymer-metal Composites (IPMCs) as Biomimetic Sensors, Actuators and Artificial Muscles - A Review, ’’ Smart Materials and Structures, 7:R15-R30.

36.

Sackmann, E. 1996. ‘‘Supported Membranes: Scientific and Practical Applications,’’ Science, 271:43-48.

37.

Singh, A. and Manjula, D. 2003. ‘‘Bacteriorhodopsin Analogs from Diphenylpolyene Chromophores,’’ Photochemistry and Photobiology , 78:503-510.

38.

Sondergaard, T.E. , Schulz, A. and Palmgren, M.G. 2004. ‘‘Energization of Transport Processes in Plants Roles of the Plasma Membrane H+-ATPase,’’ Plant Physiology, 136:2475-2482.

39.

Stevens, T.H. and Forgac, M. 1997. ‘‘Structure, Function and Regulation of Vacuolar H+-ATPases,’’ Annual Reviews in Cell Developmental Biology, 13:779-808.

40.

Trojanowicz, M. and Mulchandni, A. 2004. ‘‘Analytical Applications of Planar Bilayer Lipid Membranes,’’ Analytical and Bioanalytical Chemistry, 379:347-350.

41.

Tosteson, D. and Hoffman, J. 1960. ‘‘Regulation of Cell Volume by Active Cation Transport in High and Low Potassium Sheep Red Cells,’’ Journal of General Physiology, 44:169-194.

42.

Whittam, R. and Wheeler, K. 1970. ‘‘Transport Across Cell Membranes, ’’ Annual Review of Physiology, 32:21-60.

43.

Worsfold, O. , Toma, C. and Nishiya, T. 2004. ‘‘Development of a Novel Optical Bionanosensor ,’’ Biosensors and Bioelectronics, 19:1505-1511.