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Reconstitution of H+ATPase into Planar Phospholipid Bilayers and Its Kinetic Analysis

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Summary

The proton-translocating ATPase of the thermophilic bacterium PS3 (TFoF1) was incorporated into planar phospholipid bilayers, and its electrogenicity and kinetic characteristics were examined. A short-circuit current of up to 1 nA/cm2 was generated upon the addition of ATP. The generation of the electric current was progressively suppressed by inhibitors of TF1. From the reversal potential, the electrogenicity of TFoF1 was indicated to be some 180 mV. The relationship between the electric current induced by ATP and the concentration of ATP revealed that the magnitude of the electric current followed simple Michaelis-Menten type kinetics and the Km was found to be 0.14 mM under the conditions studied. Furthermore, there was no apparent dependence of the Km on externally applied membrane potential. These results suggest that the voltage dependence resides in some steps that defines the apparent Vmax rather than Km in the reaction cycle.

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References

  1. Boyer, P.D., Cross, R.L., and Momsen, W. (1973) Proc. Natl. Acad. Sci. 70, 2873–2839

    Article  Google Scholar 

  2. Grubmeyer, C., and Penefsky, H.S. (1981) J. Biol. Chem. 256, 3718–3727

    PubMed  CAS  Google Scholar 

  3. Takabe, T., and Hammes, G.G. (1981) Biochemistry 20, 6859–6864

    Article  PubMed  CAS  Google Scholar 

  4. Wong, S.Y., Matsuno-Yagi, A., and Hatefi, Y. (1984) Biochemistry 23, 5004–5009

    Article  PubMed  CAS  Google Scholar 

  5. Kagawa, Y., and Sone, N. (1979) Methods Enzymol. 55, 364–372

    Article  PubMed  CAS  Google Scholar 

  6. Hirata, H., Ohno, K., Sone, N., Kagawa, Y., and Hamamoto, T. (1986) J. Biol. Chem. 261, 9839–9843

    PubMed  CAS  Google Scholar 

  7. Muneyuki, E., Kagawa, Y., and Hirata, H. (1989) J. Biol. Chem. 264, 6092–6096

    PubMed  CAS  Google Scholar 

  8. Montai, M., and Mueller, P. (1972) Proc. Natl. Acad. Sci. USA 69, 3561–3566

    Article  Google Scholar 

  9. Cross, R.L., Grubmeyer, C., and Penefsky, H.S. (1982) J. Biol. Chem. 257, 12101–12105

    PubMed  CAS  Google Scholar 

  10. Gresser, M.J., Myers, J.A., and Boyer, P.D. (1982) J. Biol. Chem. 257, 12030–12038

    PubMed  CAS  Google Scholar 

  11. Rogner, M., and Gräber, P. (1986) Eur. J. Biochem. 159, 255–261

    Article  PubMed  CAS  Google Scholar 

  12. Muneyuki, E., and Hirata, H. (1988) FEBS Lett. 234, 455–458

    Article  PubMed  CAS  Google Scholar 

  13. Hatefi, Y., Yagi, T., Phelps, D.C., Wong, S-Y., Vik, S.B., and Galante, Y.M. (1982) Proc. Natl. Acad. Sci. USA 79, 1756–1760

    Article  PubMed  CAS  Google Scholar 

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© 1990 Plenum Press, New York

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Hirata, H., Muneyuki, E. (1990). Reconstitution of H+ATPase into Planar Phospholipid Bilayers and Its Kinetic Analysis. In: Kim, C.H., Ozawa, T. (eds) Bioenergetics. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5835-0_22

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  • DOI: https://doi.org/10.1007/978-1-4684-5835-0_22

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4684-5837-4

  • Online ISBN: 978-1-4684-5835-0

  • eBook Packages: Springer Book Archive

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