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. 1983 Oct;156(1):81–88. doi: 10.1128/jb.156.1.81-88.1983

2-Oxoacid dehydrogenase complexes of Escherichia coli: cellular amounts and patterns of synthesis.

M W Smith, F C Neidhardt
PMCID: PMC215053  PMID: 6311808

Abstract

The oxidative decarboxylations of pyruvate and 2-oxoglutarate in Escherichia coli are carried out by two large, multienzyme complexes: pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase. The enzyme complexes each contain three subunits: two are unique to the individual complexes, the third is shared between them. Resolution of the polypeptide subunits on two-dimensional gels allowed quantitative analysis of their cellular levels and patterns of synthesis in growing cells. Cells growing in glucose-salts medium were found to contain roughly 85 to 136 pyruvate dehydrogenase complexes and 73 2-oxoglutarate complexes. Lipoamide dehydrogenase, the subunit shared by the two complexes, was found to be in significant excess of its stoichiometric demand in the two enzyme complexes under most growth conditions. The subunits unique to each of the complexes were coordinately regulated over a wide variety of growth conditions and a broad range of expression. The two complexes responded to different, but partially overlapping, regulatory signals. Most importantly, the shared subunit was actively regulated to accommodate its demand in both enzymes. These results are discussed with regard to possible mechanisms of regulation of the enzyme complexes in general and of the shared subunit specifically.

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Selected References

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

  1. Amarasingham C. R., Davis B. D. Regulation of alpha-ketoglutarate dehydrogenase formation in Escherichia coli. J Biol Chem. 1965 Sep;240(9):3664–3668. [PubMed] [Google Scholar]
  2. Bleile D. M., Munk P., Oliver R. M., Reed L. J. Subunit structure of dihydrolipoyl transacetylase component of pyruvate dehydrogenase complex from Escherichia coli. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4385–4389. doi: 10.1073/pnas.76.9.4385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bloch P. L., Phillips T. A., Neidhardt F. C. Protein identifications of O'Farrell two-dimensional gels: locations of 81 Escherichia coli proteins. J Bacteriol. 1980 Mar;141(3):1409–1420. doi: 10.1128/jb.141.3.1409-1420.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blumenthal R. M., Reeh S., Pedersen S. Regulation of transcription factor rho and the alpha subunit of RNA polymerase in Escherichia coli B/r. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2285–2288. doi: 10.1073/pnas.73.7.2285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Creaghan I. T., Guest J. R. Amber mutants of the -ketoglutarate dehydrogenase gene of Escherichia coli K12. J Gen Microbiol. 1972 Jul;71(2):207–220. doi: 10.1099/00221287-71-2-207. [DOI] [PubMed] [Google Scholar]
  7. Derosier D. J., Oliver R. M., Reed L. J. Crystallization and preliminary structural analysis of dihydrolipoyl transsuccinylase, the core of the 2-oxoglutarate dehydrogenase complex. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1135–1137. doi: 10.1073/pnas.68.6.1135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dietrich J., Henning U. Regulation of pyruvate dehydrogenase complex synthesis in Escherichia coli K 12. Identification of the inducing metabolite. Eur J Biochem. 1970 Jun;14(2):258–269. doi: 10.1111/j.1432-1033.1970.tb00285.x. [DOI] [PubMed] [Google Scholar]
  9. Gray C. T., Wimpenny J. W., Mossman M. R. Regulation of metabolism in facultative bacteria. II. Effects of aerobiosis, anaerobiosis and nutrition on the formation of Krebs cycle enzymes in Escherichia coli. Biochim Biophys Acta. 1966 Mar 28;117(1):33–41. doi: 10.1016/0304-4165(66)90149-8. [DOI] [PubMed] [Google Scholar]
  10. Guest J. R., Cole S. T., Jeyaseelan K. Organization and expression of the pyruvate dehydrogenase complex genes of Escherichia coli K12. J Gen Microbiol. 1981 Nov;127(1):65–79. doi: 10.1099/00221287-127-1-65. [DOI] [PubMed] [Google Scholar]
  11. Guest J. R., Creaghan I. T. Gene-protein relationships of the alpha-keto acid dehydrogenase complexes of Escherichia coli K12: isolation and characterization of lipoamide dehydrogenase mutants. J Gen Microbiol. 1973 Mar;75(1):197–210. doi: 10.1099/00221287-75-1-197. [DOI] [PubMed] [Google Scholar]
  12. Guest J. R. Gene-protein relationships of the alpha-keto acid dehydrogenase complexes of Escherichia coli K12: Chromosomal location of the lipoamide dehydrogenase gene. J Gen Microbiol. 1974 Feb;80(2):523–532. doi: 10.1099/00221287-80-2-523. [DOI] [PubMed] [Google Scholar]
  13. Guest J. R., Stephens P. E. Molecular cloning of the pyruvate dehydrogenase complex genes of Escherichia coli. J Gen Microbiol. 1980 Dec;121(2):277–292. doi: 10.1099/00221287-121-2-277. [DOI] [PubMed] [Google Scholar]
  14. Hirshfield I. N., Bloch P. L., Van Bogelen R. A., Neidhardt F. C. Multiple forms of lysyl-transfer ribonucleic acid synthetase in Escherichia coli. J Bacteriol. 1981 Apr;146(1):345–351. doi: 10.1128/jb.146.1.345-351.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Langley D., Guest J. R. Biochemical genetics of the alpha-keto acid dehydrogenase complexes of Escherichia coli K12: genetic characterization and regulatory properties of deletion mutants. J Gen Microbiol. 1978 May;106(1):103–117. doi: 10.1099/00221287-106-1-103. [DOI] [PubMed] [Google Scholar]
  16. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Pedersen S., Bloch P. L., Reeh S., Neidhardt F. C. Patterns of protein synthesis in E. coli: a catalog of the amount of 140 individual proteins at different growth rates. Cell. 1978 May;14(1):179–190. doi: 10.1016/0092-8674(78)90312-4. [DOI] [PubMed] [Google Scholar]
  19. Phillips T. A., Bloch P. L., Neidhardt F. C. Protein identifications on O'Farrell two-dimensional gels: locations of 55 additional Escherichia coli proteins. J Bacteriol. 1980 Dec;144(3):1024–1033. doi: 10.1128/jb.144.3.1024-1033.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith M. W., Neidhardt F. C. Proteins induced by aerobiosis in Escherichia coli. J Bacteriol. 1983 Apr;154(1):344–350. doi: 10.1128/jb.154.1.344-350.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith M. W., Neidhardt F. C. Proteins induced by anaerobiosis in Escherichia coli. J Bacteriol. 1983 Apr;154(1):336–343. doi: 10.1128/jb.154.1.336-343.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Spencer M. E., Guest J. R. Molecular cloning of four tricarboxylic acid cyclic genes of Escherichia coli. J Bacteriol. 1982 Aug;151(2):542–552. doi: 10.1128/jb.151.2.542-552.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wanner B. L., Kodaira R., Neidhardt F. C. Physiological regulation of a decontrolled lac operon. J Bacteriol. 1977 Apr;130(1):212–222. doi: 10.1128/jb.130.1.212-222.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wanner B. L., McSharry R. Phosphate-controlled gene expression in Escherichia coli K12 using Mudl-directed lacZ fusions. J Mol Biol. 1982 Jul 5;158(3):347–363. doi: 10.1016/0022-2836(82)90202-9. [DOI] [PubMed] [Google Scholar]
  25. Yamamoto I., Ishimoto M. Effect of nitrate reduction on the enzyme levels in carbon metabolism in Escherichia coli. J Biochem. 1975 Aug;78(2):307–315. doi: 10.1093/oxfordjournals.jbchem.a130909. [DOI] [PubMed] [Google Scholar]

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