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. 1981 Apr;78(4):2140–2144. doi: 10.1073/pnas.78.4.2140

Capacity for alternating sites cooperativity in catalysis by succinyl-coenzyme A synthetase.

W T Wolodko, M D O'Connor, W A Bridger
PMCID: PMC319299  PMID: 7017725

Abstract

Succinyl-coenzyme A synthetase [succinate:CoA ligase (ADP-forming), EC 6.2.1.5] of Escherichia coli in an alpha 2 beta 2 tetramer. A histidyl residue in the alpha subunit is phosphorylated as a catalytic intermediate. It has been suggested [Bild, G. S., Janson, C. & Boyer, P. D. (1980) J. Biol. Chem. 255, 8109--8115] that the mechanism of action of this enzyme involves intersubunit cooperativity in which attachment of substrates at one of the two active sites promotes catalytic events at the other. This scheme would require that the two active sites, although otherwise equivalent, should act alternately. We have prepared a hybrid enzyme species that contains one 35S-labeled alpha subunit (dephosphorylated), one nonradioactive alpha subunit (phosphorylated), and two beta subunits per tetrameric molecule. With the aid of a selective chromatographic procedure for the isolation of peptides that contain phosphohistidyl residues, we have shown that each of the alpha subunits undergoes phosphorylation when the hybrid enzyme is exposed briefly to substrates. This result demonstrates that the two active sites are capable of alternate activity and lends support to the concept of alternating sites cooperativity. The half-of-the-sites phosphorylation that occurs with this enzyme is not a consequence of permanent asymmetry or other lack of equivalence of the two alpha subunits.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bild G. S., Janson C. A., Boyer P. D. Subunit interaction during catalysis. ATP modulation of catalytic steps in the succinyl-CoA synthetase reaction. J Biol Chem. 1980 Sep 10;255(17):8109–8115. [PubMed] [Google Scholar]
  2. Bridger W. A. Evidence for two types of subunits in succinyl coenzyme A synthetase. Biochem Biophys Res Commun. 1971 Mar 5;42(5):948–954. doi: 10.1016/0006-291x(71)90522-5. [DOI] [PubMed] [Google Scholar]
  3. Chan W. W. Some experimental approaches for studying subunit interactions in enzymes. Can J Biochem. 1976 Jun;54(6):521–528. doi: 10.1139/o76-076. [DOI] [PubMed] [Google Scholar]
  4. Collier G. E., Nishimura J. S. Affinity labeling of succinyl-CoA synthetase from porcine heart and Escherichia coli with oxidized coenzyme A disulfide. J Biol Chem. 1978 Jul 25;253(14):4938–4943. [PubMed] [Google Scholar]
  5. Evans P. R., Hudson P. J. Structure and control of phosphofructokinase from Bacillus stearothermophilus. Nature. 1979 Jun 7;279(5713):500–504. doi: 10.1038/279500a0. [DOI] [PubMed] [Google Scholar]
  6. Fletterick R. J., Madsen N. B. The structures and related functions of phosphorylase a. Annu Rev Biochem. 1980;49:31–61. doi: 10.1146/annurev.bi.49.070180.000335. [DOI] [PubMed] [Google Scholar]
  7. Krebs A., Bridger W. A. Some physical parameters of succinyl-coenzyme A synthetase of Escherichia coli. Can J Biochem. 1974 Jul;52(7):594–598. doi: 10.1139/o74-086. [DOI] [PubMed] [Google Scholar]
  8. Leitzmann C., Wu J. Y., Boyer P. D. Subunits, composition, and related properties of succinyl coenzyme A synthetase. Biochemistry. 1970 May 26;9(11):2338–2346. doi: 10.1021/bi00813a018. [DOI] [PubMed] [Google Scholar]
  9. Moffet F. J., Wang T., Bridger W. A. Succinyl coenzyme A synthetase of Escherichia coli. Effects of phosphoenzyme formation and of substrate binding on the reactivity and stability of the enzyme. J Biol Chem. 1972 Dec 25;247(24):8139–8144. [PubMed] [Google Scholar]
  10. Pearson P. H., Bridger W. A. Catalysis of a step of the overall reaction by the alpha subunit of Escherichia coli succinyl coenzyme A synthetase. J Biol Chem. 1975 Nov 10;250(21):8524–8529. [PubMed] [Google Scholar]
  11. Pearson P. H., Bridger W. A. Isolation of the alpha and beta subunits of Escherichia coli succinyl coenzyme A synthetase and their recombination into active enzyme. J Biol Chem. 1975 Jun 25;250(12):4451–4455. [PubMed] [Google Scholar]
  12. Wang T., Jurásek L., Bridger W. A. Succinyl coenzyme A synthetase of Escherichia coli. Sequence of a peptide containing the active-site phosphohistidine residue. Biochemistry. 1972 May 23;11(11):2067–2070. doi: 10.1021/bi00761a011. [DOI] [PubMed] [Google Scholar]
  13. Wolodko W. T., Brownie E. R., Bridger W. A. Subunits of succinyl-coenzyme A synthetase: coordination of production in Escherichia coli and discovery of a factor that precludes refolding. J Bacteriol. 1980 Jul;143(1):231–237. doi: 10.1128/jb.143.1.231-237.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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