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.
Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP Synthetase Complexes / chemistry
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ATP Synthetase Complexes / genetics
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ATP Synthetase Complexes / metabolism*
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Bacterial Proton-Translocating ATPases / chemistry
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Bacterial Proton-Translocating ATPases / genetics
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Bacterial Proton-Translocating ATPases / metabolism*
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Benzophenones / chemistry
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Benzophenones / pharmacology
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Binding Sites / genetics
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Cross-Linking Reagents / chemistry
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Cross-Linking Reagents / pharmacology
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Cysteine / chemistry
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Cysteine / genetics
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Cysteine / metabolism
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Disulfides / chemistry
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Disulfides / pharmacology
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Escherichia coli / enzymology*
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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Immunoblotting
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Maleimides / chemistry
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Maleimides / pharmacology
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Models, Molecular
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Mutation
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Protein Binding / drug effects
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Protein Multimerization
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Protein Structure, Tertiary
Substances
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Benzophenones
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Cross-Linking Reagents
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Disulfides
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Escherichia coli Proteins
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Maleimides
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benzophenone-4-maleimide
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ATP Synthetase Complexes
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ATP synthase b subunit, E coli
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Bacterial Proton-Translocating ATPases
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atpB protein, E coli
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Cysteine