Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1985 Feb 1;225(3):731–735. doi: 10.1042/bj2250731

Resolution and reconstitution of bovine kidney branched-chain 2-oxo acid dehydrogenase complex.

K G Cook, A P Bradford, S J Yeaman
PMCID: PMC1144650  PMID: 3977856

Abstract

Branched-chain 2-oxo acid dehydrogenase complex was resolved into component E1 and E2-kinase subcomplex by gel filtration in the presence of 1 M-NaC1. Essentially all the original activity of the complex can be regained after reconstitution of the component enzymes, reassociation being a rapid process. The specific activities of E1 and E2 were 25.1 and 19.0 units/mg respectively. Non-phosphorylated active E1 has an approx. 6-fold higher affinity for E2 than does phosphorylated E1. The components of the branched-chain 2-oxo acid dehydrogenase complex do not crossreact with the respective components from the pyruvate dehydrogenase complex. The significance of these results and of the tight association of the kinase with E2 are discussed.

Full text

PDF
731

Images in this article

Selected References

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

  1. Barrera C. R., Namihira G., Hamilton L., Munk P., Eley M. H., Linn T. C., Reed L. J. -Keto acid dehydrogenase complexes. XVI. Studies on the subunit structure of the pyruvate dehydrogenase complexes from bovine kidney and heart. Arch Biochem Biophys. 1972 Feb;148(2):343–358. doi: 10.1016/0003-9861(72)90152-x. [DOI] [PubMed] [Google Scholar]
  2. Brandt D. R., Roche T. E., Pratt M. L. Heterogeneity of binding sites for the pyruvate dehydrogenase component on the dihydrolipoyl transacetylase core of bovine kidney pyruvate dehydrogenase complex. Biochemistry. 1983 Jun 7;22(12):2958–2965. doi: 10.1021/bi00281a027. [DOI] [PubMed] [Google Scholar]
  3. Brandt D. R., Roche T. E. Specificity of the pyruvate dehydrogenase kinase for pyruvate dehydrogenase component bound to the surface of the kidney pyruvate dehydrogenase complex and evidence for intracore migration of pyruvate dehydrogenase component. Biochemistry. 1983 Jun 7;22(12):2966–2971. doi: 10.1021/bi00281a028. [DOI] [PubMed] [Google Scholar]
  4. Cate R. L., Roche T. E., Davis L. C. Rapid intersite transfer of acetyl groups and movement of pyruvate dehydrogenase component in the kidney pyruvate dehydrogenase complex. J Biol Chem. 1980 Aug 25;255(16):7556–7562. [PubMed] [Google Scholar]
  5. Cook K. G., Lawson R., Yeaman S. J., Aitken A. Amino acid sequence at the major phosphorylation site on bovine kidney branched-chain 2-oxoacid dehydrogenase complex. FEBS Lett. 1983 Nov 28;164(1):47–50. doi: 10.1016/0014-5793(83)80016-7. [DOI] [PubMed] [Google Scholar]
  6. Fatania H. R., Lau K. S., Randle P. J. Inactivation of purified ox kidney branched-chain 2-oxoacid dehydrogenase complex by phosphorylation. FEBS Lett. 1981 Sep 28;132(2):285–288. doi: 10.1016/0014-5793(81)81180-5. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  8. Lawson R., Cook K. G., Yeaman S. J. Rapid purification of bovine kidney branched-chain 2-oxoacid dehydrogenase complex containing endogenous kinase activity. FEBS Lett. 1983 Jun 27;157(1):54–58. doi: 10.1016/0014-5793(83)81115-6. [DOI] [PubMed] [Google Scholar]
  9. Linn T. C., Pelley J. W., Pettit F. H., Hucho F., Randall D. D., Reed L. J. -Keto acid dehydrogenase complexes. XV. Purification and properties of the component enzymes of the pyruvate dehydrogenase complexes from bovine kidney and heart. Arch Biochem Biophys. 1972 Feb;148(2):327–342. doi: 10.1016/0003-9861(72)90151-8. [DOI] [PubMed] [Google Scholar]
  10. Odessey R. Purification of rat kidney branched-chain oxo acid dehydrogenase complex with endogenous kinase activity. Biochem J. 1982 Apr 15;204(1):353–356. doi: 10.1042/bj2040353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pettit F. H., Yeaman S. J., Reed L. J. Purification and characterization of branched chain alpha-keto acid dehydrogenase complex of bovine kidney. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4881–4885. doi: 10.1073/pnas.75.10.4881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pratt M. L., Roche T. E., Dyer D. W., Cate R. L. Enhanced dissociation of pyruvate dehydrogenase from the pyruvate dehydrogenase complex following phosphorylation and regulatory implications. Biochem Biophys Res Commun. 1979 Nov 14;91(1):289–296. doi: 10.1016/0006-291x(79)90616-8. [DOI] [PubMed] [Google Scholar]
  13. Stanley C. J., Perham R. N. Purification of 2-oxo acid dehydrogenase multienzyme complexes from ox heart by a new method. Biochem J. 1980 Oct 1;191(1):147–154. doi: 10.1042/bj1910147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wu T. L., Reed L. J. Subunit binding in the pyruvate dehydrogenase complex from bovine kidney and heart. Biochemistry. 1984 Jan 17;23(2):221–226. doi: 10.1021/bi00297a008. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES