Regular paper
The position of the ATP binding site on the (Ca2+ + Mg2+)-ATPase

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Abstract

We present a convenient method to calculate the efficiency of fluorescence energy transfer in two-dimensional membrane systems. We apply it to the analysis of energy transfer between phospholipid molecules labelled with fluorescein and rhodamine groups, and of energy transfer in reconstituted membranes containing (Ca2+ + Mg2+)-ATPase purified from sarcoplasmic reticulum, with the ATPase labelled at the ATP binding site with fluorescein as donor, and rhodamine-labelled lipid as acceptor. The ATP binding site is found to be distant from the plane of the lipid/water interface of the membrane. It is suggested that the ATPase is present in the membrane as a dimer, with the two ATP binding sites in the dimer being close to the protein/protein interface. Addition of vanadate causes no change in quenching, suggesting that the ATP binding site does not move significantly with respect to the lipid/water interface in the E1-E2 conformational transition of the ATPase.

References (46)

  • N. Ikemoto et al.

    J. Biol. Chem.

    (1981)
  • L. Dux et al.

    J. Biol. Chem.

    (1983)
  • N. Ikemoto et al.

    J. Biol. Chem.

    (1984)
  • D.W. Martin et al.

    FEBS Lett.

    (1984)
  • J.P. Andersen et al.

    J. Biol. Chem.

    (1985)
  • S.R. Highsmith et al.

    J. Biol. Chem.

    (1983)
  • S. Highsmith

    Biochem. Biophys. Res. Commun.

    (1984)
  • S. Highsmith et al.

    J. Biol. Chem.

    (1984)
  • T.L. Scott

    J. Biol. Chem.

    (1985)
  • U. Pick et al.

    Biochim. Biophys. Acta

    (1980)
  • O.H. Lowry et al.

    J. Biol. Chem.

    (1951)
  • M. Kasahara et al.

    J. Biol. Chem.

    (1977)
  • P.K. Wolber et al.

    Biophys. J.

    (1979)
  • B. Snyder et al.

    Biophys. J.

    (1982)
  • C. Gutierrez-Merino

    Biophys. Chem.

    (1981)
  • C. Gutierrez-Merino

    Biophys. Chem.

    (1981)
  • A.G. Lee

    FEBS Lett.

    (1983)
  • C.D. Stubbs et al.

    Biochim. Biophys. Acta

    (1984)
  • K.A. Taylor et al.

    J. Mol. Biol.

    (1986)
  • L. Hymel et al.

    J. Biol. Chem.

    (1984)
  • A.M. Mata et al.

    Biochem. Biophys. Res. Commun.

    (1985)
  • U. Pick

    J. Biol. Chem.

    (1982)
  • M. Tada et al.

    Physiol. Rev.

    (1978)
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