Disulfide bond formation between the COOH-terminal domain of the β subunits and the γ subunits and ∈ subunits of the Escherichia coli F1-atpase: structural implications and functional consequences

Aggeler, Robert, Haughton, Margaret A. and Capaldi, Roderick A. (1995) Disulfide bond formation between the COOH-terminal domain of the β subunits and the γ subunits and ∈ subunits of the Escherichia coli F1-atpase: structural implications and functional consequences. Journal of Biological Chemistry, 270 16: 9185-9191. doi:10.1074/jbc.270.16.9185

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Author Aggeler, Robert
Haughton, Margaret A.
Capaldi, Roderick A.
Title Disulfide bond formation between the COOH-terminal domain of the β subunits and the γ subunits and ∈ subunits of the Escherichia coli F1-atpase: structural implications and functional consequences
Formatted title
Disulfide bond formation between the COOH-terminal domain of the β subunits and the γ subunits and ∈ subunits of the Escherichia coli F1-atpase: structural implications and functional consequences
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
1083-351X
Publication date 1995-04
Sub-type Article (original research)
DOI 10.1074/jbc.270.16.9185
Open Access Status File (Publisher version)
Volume 270
Issue 16
Start page 9185
End page 9191
Total pages 7
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Formatted abstract
A set of mutants of the Escherichia coli F1F0-type ATPase has been generated by site-directed mutagenesis as follows: βE381C, βS383C, βE381C/∈S108C, and βS383C/∈S108C. Treatment of ECFisolated from any of these mutants with CuCl2 induces disulfide bond formation. For the single mutants, βE381C and βS383C, a disulfide bond is formed in essentially 100% yield between a β subunit and the γ subunit, probably at Cys87 based on the recent structure determination of F1(Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628). In the double mutants, two disulfide bonds are formed, again in essentially full yield, one between β and γ, the other between a β and the ∈ subunit via Cys108. The same two cross-links are produced with CuCl2 treatment of ECF1F0 isolated from either of the double mutants. These results show that the parts of γ around residue 87 (a short α-helix) and the ∈ subunit interact with different β subunits.

The yield of covalent linkage of β to γ is nucleotide dependent and highest in ATP and much lower with ADP in catalytic sites. The yield of covalent linkage of β to ∈ is also nucleotide dependent but in this case is highest in ADP and much lower in ATP. Disulfide bond formation between either β and γ, or β and ∈ inhibits the ATPase activity of the enzyme in proportion to the yield of the cross-linked product. Chemical modification of the Cys at either position 381 or 383 of the β subunit inhibits ATPase activity in a manner that appears to be dependent on the size of the modifying reagent. These results are as expected if movements of the catalytic site-containing β subunits relative to the γ and ∈ subunits are an essential part of the cooperativity of the enzyme.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads
 
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