Interactions between subunits a and b in the rotary ATP synthase as determined by cross-linking

FEBS Lett. 2013 Apr 2;587(7):892-7. doi: 10.1016/j.febslet.2013.02.012. Epub 2013 Feb 14.

Abstract

The interaction of the membrane traversing stator subunits a and b of the rotary ATP synthase was probed by substitution of a single Cys into each subunit with subsequent Cu(2+) catalyzed cross-linking. Extensive interaction between the transmembrane (TM) region of one b subunit and TM2 of subunit a was indicated by cross-linking with 6 Cys pairs introduced into these regions. Additional disulfide cross-linking was observed between the N-terminus of subunit b and the periplasmic loop connecting TM4 and TM5 of subunit a. Finally, benzophenone-4-maleimide derivatized Cys in the 2-3 periplasmic loop of subunit a were shown to cross-link with the periplasmic N-terminal region of subunit b. These experiments help to define the juxtaposition of subunits b and a in the ATP synthase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Synthetase Complexes / chemistry
  • ATP Synthetase Complexes / genetics
  • ATP Synthetase Complexes / metabolism*
  • Bacterial Proton-Translocating ATPases / chemistry
  • Bacterial Proton-Translocating ATPases / genetics
  • Bacterial Proton-Translocating ATPases / metabolism*
  • Benzophenones / chemistry
  • Benzophenones / pharmacology
  • Binding Sites / genetics
  • Cross-Linking Reagents / chemistry
  • Cross-Linking Reagents / pharmacology
  • Cysteine / chemistry
  • Cysteine / genetics
  • Cysteine / metabolism
  • Disulfides / chemistry
  • Disulfides / pharmacology
  • Escherichia coli / enzymology*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Immunoblotting
  • Maleimides / chemistry
  • Maleimides / pharmacology
  • Models, Molecular
  • Mutation
  • Protein Binding / drug effects
  • Protein Multimerization
  • Protein Structure, Tertiary

Substances

  • Benzophenones
  • Cross-Linking Reagents
  • Disulfides
  • Escherichia coli Proteins
  • Maleimides
  • benzophenone-4-maleimide
  • ATP Synthetase Complexes
  • ATP synthase b subunit, E coli
  • Bacterial Proton-Translocating ATPases
  • atpB protein, E coli
  • Cysteine