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. 1979 Mar;76(3):1213–1217. doi: 10.1073/pnas.76.3.1213

Requirements of acetyl phosphate for the binding protein-dependent transport systems in Escherichia coli.

J S Hong, A G Hunt, P S Masters, M A Lieberman
PMCID: PMC383220  PMID: 375230

Abstract

In Escherichia coli, acetyl phosphate can be formed from acetyl-CoA via the phosphotransacetylase (phosphate acetyltransferase; acetyl-CoA:orthophosphate acetyltransferase, EC 2.3.1.8) reaction and from acetate (plus ATP) via the acetate kinase (ATP:acetate phosphotransferase, EC 2.7.2.1) reaction. By restricting acetyl phosphate formation to the phosphotransacetylase reaction alone, through the use of metabolic inhibitors, we were able to show that, with pyruvate as a source of energy, mutants defective in phosphotransacetylase are unable to transport glutamine, histidine, and methionine. However, with the same energy source, mutants defective in acetate kinase are normal in the transport of these amino acids. The inability of the phosphotransacetylase mutants to transport is due to their presumed inability to form acetyl phosphate, because pyruvate is found to be metabolized to acetyl-CoA in these mutants. Thus acetyl phosphate has been implicated in active transport. Evidence is also presented that neither the protonmotive force nor the ecf gene product is required for the shock-sensitive transport systems.

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Selected References

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

  1. Altendorf K., Hirata H., Harold F. M. Accumulation of lipid-soluble ions and of rubidium as indicators of the electrical potential in membrane vesicles of Escherichia coli. J Biol Chem. 1975 Feb 25;250(4):1405–1412. [PubMed] [Google Scholar]
  2. Berger E. A. Different mechanisms of energy coupling for the active transport of proline and glutamine in Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1514–1518. doi: 10.1073/pnas.70.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berger E. A., Heppel L. A. Different mechanisms of energy coupling for the shock-sensitive and shock-resistant amino acid permeases of Escherichia coli. J Biol Chem. 1974 Dec 25;249(24):7747–7755. [PubMed] [Google Scholar]
  4. Brown T. D., Jones-Mortimer M. C., Kornberg H. L. The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli. J Gen Microbiol. 1977 Oct;102(2):327–336. doi: 10.1099/00221287-102-2-327. [DOI] [PubMed] [Google Scholar]
  5. Fields K. L., Luria S. E. Effects of colicins E1 and K on cellular metabolism. J Bacteriol. 1969 Jan;97(1):64–77. doi: 10.1128/jb.97.1.64-77.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gutnick D., Calvo J. M., Klopotowski T., Ames B. N. Compounds which serve as the sole source of carbon or nitrogen for Salmonella typhimurium LT-2. J Bacteriol. 1969 Oct;100(1):215–219. doi: 10.1128/jb.100.1.215-219.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hong J. S. An ecf mutation in Escherichia coli pleiotropically affecting energy coupling in active transport but not generation or maintenance of membrane potential. J Biol Chem. 1977 Dec 10;252(23):8582–8588. [PubMed] [Google Scholar]
  8. Janson C. A., Cleland W. W. The inhibition of acetate, pyruvate, and 3-phosphogylcerate kinases by chromium adenosine triphosphate. J Biol Chem. 1974 Apr 25;249(8):2567–2571. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lieberman M. A., Hong J. S. A mutant of Escherichia coli defective in the coupling of metabolic energy to active transport. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4395–4399. doi: 10.1073/pnas.71.11.4395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lieberman M. A., Hong J. S. Changes in active transport, intracellular adenosine 5'-triphosphate levels, macromolecular syntheses, and glycolysis in an energy-uncoupled mutant of Escherichia coli. J Bacteriol. 1976 Mar;125(3):1024–1031. doi: 10.1128/jb.125.3.1024-1031.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lieberman M. A., Hong J. S. Energization of osmotic shock-sensitive transport systems in Escherichia coli requires more than ATP. Arch Biochem Biophys. 1976 Jan;172(1):312–315. doi: 10.1016/0003-9861(76)90080-1. [DOI] [PubMed] [Google Scholar]
  13. Lieberman M. A., Simon M., Hong J. S. Characterization of Escherichia coli mutant incapable of maintaining a transmembrane potential. MetC ecfts mutations. J Biol Chem. 1977 Jun 25;252(12):4056–4067. [PubMed] [Google Scholar]
  14. Mitchell P. Performance and conservation of osmotic work by proton-coupled solute porter systems. J Bioenerg. 1973 Jan;4(1):63–91. doi: 10.1007/BF01516051. [DOI] [PubMed] [Google Scholar]
  15. PEARSON D. J. A SOURCE OF ERROR IN THE ASSAY OF ACETYL-COENZYME A. Biochem J. 1965 Jun;95:23C–24C. doi: 10.1042/bj0950023c. [DOI] [PubMed] [Google Scholar]
  16. Pauli G., Overath P. ato Operon: a highly inducible system for acetoacetate and butyrate degradation in Escherichia coli. Eur J Biochem. 1972 Sep 25;29(3):553–562. doi: 10.1111/j.1432-1033.1972.tb02021.x. [DOI] [PubMed] [Google Scholar]
  17. Plate C. A., Suit J. L., Jetten A. M., Luria S. E. Effects of colicin K on a mutant of Escherichia coli deficient in Ca 2+, Mg 2+-activated adenosine triphosphatase. J Biol Chem. 1974 Oct 10;249(19):6138–6143. [PubMed] [Google Scholar]
  18. Ramos S., Schuldiner S., Kaback H. R. The electrochemical gradient of protons and its relationship to active transport in Escherichia coli membrane vesicles. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1892–1896. doi: 10.1073/pnas.73.6.1892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Saier M. H., Jr, Wentzel D. L., Feucht B. U., Judice J. J. A transport system for phosphoenolpyruvate, 2-phosphoglycerate, and 3-phosphoglycerate in Salmonella typhimurium. J Biol Chem. 1975 Jul 10;250(13):5089–5096. [PubMed] [Google Scholar]
  20. Simoni R. D., Postma P. W. The energetics of bacterial active transport. Annu Rev Biochem. 1975;44:523–554. doi: 10.1146/annurev.bi.44.070175.002515. [DOI] [PubMed] [Google Scholar]
  21. Singh A. P., Bragg P. D. Energetics of galactose, proline, and glutamine transport in a cytochrome-deficient mutant of Salmonella typhimurium. J Supramol Struct. 1977;6(3):389–398. doi: 10.1002/jss.400060312. [DOI] [PubMed] [Google Scholar]
  22. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]

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