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Characterization of three-subunit chloroplast coupling factor.

Publication ,  Journal Article
Mitra, B; Hammes, GG
Published in: Biochemistry
January 12, 1988

The delta- and epsilon-polypeptides were removed from chloroplast coupling factor 1 (CF1). The resulting enzyme, CF1(-delta, epsilon), is a stable active ATPase containing only alpha-, beta-, and gamma-polypeptides. The dependence of the steady-state kinetics of ATP hydrolysis catalyzed by CF1(-delta, epsilon) on the concentrations of ATP and ADP was found to be essentially the same as by activated CF1. Nucleotide binding studies with CF1(-delta, epsilon) revealed three binding sites: a nondissociable ADP site (site 1), a tight MgATP binding site (site 2), and a site that binds ADP and ATP with a dissociation constant in the micromolar range (site 3). Similar results have been obtained with CF1. For both CF1 and CF1(-delta, epsilon), the binding of MgATP at site 2 is tight only in the presence of Mg2+. Fluorescence resonance energy transfer was used to map distances between the gamma-sulfhydryl ("dark" site) and gamma-disulfide and between the gamma-sulfhydryl and the three nucleotide sites. These distances are within 5% of the corresponding distances on CF1. These results indicate that removal of the delta- and epsilon-polypeptides from CF1 does not cause significant changes in the structure, kinetics, and nucleotide binding sites of the enzyme.

Duke Scholars

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

January 12, 1988

Volume

27

Issue

1

Start / End Page

245 / 250

Location

United States

Related Subject Headings

  • Proton-Translocating ATPases
  • Protein Binding
  • Plants
  • Macromolecular Substances
  • Kinetics
  • Biochemistry & Molecular Biology
  • Binding Sites
  • Adenosine Triphosphate
  • Adenosine Diphosphate
  • 1101 Medical Biochemistry and Metabolomics
 

Citation

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Mitra, B., & Hammes, G. G. (1988). Characterization of three-subunit chloroplast coupling factor. Biochemistry, 27(1), 245–250. https://doi.org/10.1021/bi00401a037
Mitra, B., and G. G. Hammes. “Characterization of three-subunit chloroplast coupling factor.Biochemistry 27, no. 1 (January 12, 1988): 245–50. https://doi.org/10.1021/bi00401a037.
Mitra B, Hammes GG. Characterization of three-subunit chloroplast coupling factor. Biochemistry. 1988 Jan 12;27(1):245–50.
Mitra, B., and G. G. Hammes. “Characterization of three-subunit chloroplast coupling factor.Biochemistry, vol. 27, no. 1, Jan. 1988, pp. 245–50. Pubmed, doi:10.1021/bi00401a037.
Mitra B, Hammes GG. Characterization of three-subunit chloroplast coupling factor. Biochemistry. 1988 Jan 12;27(1):245–250.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

January 12, 1988

Volume

27

Issue

1

Start / End Page

245 / 250

Location

United States

Related Subject Headings

  • Proton-Translocating ATPases
  • Protein Binding
  • Plants
  • Macromolecular Substances
  • Kinetics
  • Biochemistry & Molecular Biology
  • Binding Sites
  • Adenosine Triphosphate
  • Adenosine Diphosphate
  • 1101 Medical Biochemistry and Metabolomics