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. 1972 Nov;69(11):3336–3340. doi: 10.1073/pnas.69.11.3336

Mutants of Salmonella typhimurium and Escherichia coli Pleiotropically Defective in Active Transport*

Jen-Shiang Hong 1,†, H R Kaback 1
PMCID: PMC389766  PMID: 4343963

Abstract

Two classes of mutants isolated from E. coli and Salmonella typhimurium are altered in respiration-coupled active transport, as studied in whole cells and/or isolated membrane vesicles. Mutant cells defective in D-lactate dehydrogenase (dld) transport amino acids and lactose normally. Membrane vesicles prepared from these mutants do not exhibit D-lactate-dependent transport, D-lactate oxidation, or D-lactate: dichlorophenolindophenol reductase activity. However, succinate-dependent transport is markedly enhanced in these mutants, without a corresponding increase in succinic dehydrogenase activity.

The second class of mutants is defective in the coupling of electron transfer to active transport. Whole cells and membrane vesicles prepared from these etc mutants exhibit markedly reduced ability to transport amino acids, despite the ability of the vesicles to oxidize D-lactate, succinate, and NADH. Vectorial phosphorylation of α-methylglucoside by these mutants is normal. Electrontransfer coupling mutants are similar phenotypically to mutants uncoupled for oxidative phosphorylation (uncA), but have normal ATPase activity. Moreover, uncA mutants catalyze active transport as well as does the wild type. These experiments indicate that the ETC component is essential for the coupling of respiratory energy to active transport, and provide further evidence that the generation or utilization of ATP is not involved in these transport mechanisms.

Keywords: energy-transfer coupling mutants, D-lactate dehydrogenase mutants, membrane vesicles, active transport

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Selected References

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

  1. Barnes E. M., Jr, Kaback H. R. Beta-galactoside transport in bacterial membrane preparations: energy coupling via membrane-bounded D-lactic dehydrogenase. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1190–1198. doi: 10.1073/pnas.66.4.1190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnes E. M., Jr, Kaback H. R. Mechanisms of active transport in isolated membrane vesicles. I. The site of energy coupling between D-lactic dehydrogenase and beta-galactoside transport in Escherichia coli membrane vesicles. J Biol Chem. 1971 Sep 10;246(17):5518–5522. [PubMed] [Google Scholar]
  3. Butlin J. D., Cox G. B., Gibson F. Oxidative phosphorylation in Escherichia coli K12. Mutations affecting magnesium ion- or calcium ion-stimulated adenosine triphosphatase. Biochem J. 1971 Aug;124(1):75–81. doi: 10.1042/bj1240075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dietz G. W. Dehydrogenase activity involved in the uptake of glucose 6-phosphate by a bacterial membrane system. J Biol Chem. 1972 Jul 25;247(14):4561–4565. [PubMed] [Google Scholar]
  5. 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]
  6. Harold F. M. Conservation and transformation of energy by bacterial membranes. Bacteriol Rev. 1972 Jun;36(2):172–230. doi: 10.1128/br.36.2.172-230.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hong J. S., Ames B. N. Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3158–3162. doi: 10.1073/pnas.68.12.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kaback H. R., Barnes E. M., Jr Mechanisms of active transport in isolated membrane vesicles. II. The mechanism of energy coupling between D-lactic dehydrogenase and beta-galactoside transport in membrane preparations from Escherichia coli. J Biol Chem. 1971 Sep 10;246(17):5523–5531. [PubMed] [Google Scholar]
  9. Kaback H. R., Milner L. S. Relationship of a membrane-bound D-(-)-lactic dehydrogenase to amino acid transport in isolated bacterial membrane preparations. Proc Natl Acad Sci U S A. 1970 Jul;66(3):1008–1015. doi: 10.1073/pnas.66.3.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaback H. R. Transport across isolated bacterial cytoplasmic membranes. Biochim Biophys Acta. 1972 Aug 4;265(3):367–416. doi: 10.1016/0304-4157(72)90014-7. [DOI] [PubMed] [Google Scholar]
  11. Kaback H. R. Transport. Annu Rev Biochem. 1970;39:561–598. doi: 10.1146/annurev.bi.39.070170.003021. [DOI] [PubMed] [Google Scholar]
  12. Kerwar G. K., Gordon A. S., Kaback H. R. Mechanisms of active transport in isolated membrane vesicles. IV. Galactose transport by isolated membrane vesicles from Escherichia coli. J Biol Chem. 1972 Jan 10;247(1):291–297. [PubMed] [Google Scholar]
  13. Konings W. N., Barnes E. M., Jr, Kaback H. R. Mechanisms of active transport in isolated membrane vesicles. 2. The coupling of reduced phenazine methosulfate to the concentrative uptake of beta-galactosides and amino acids. J Biol Chem. 1971 Oct 10;246(19):5857–5861. [PubMed] [Google Scholar]
  14. 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]
  15. Mitchell P. Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. Biol Rev Camb Philos Soc. 1966 Aug;41(3):445–502. doi: 10.1111/j.1469-185x.1966.tb01501.x. [DOI] [PubMed] [Google Scholar]
  16. Mitchell P. Proton-translocation phosphorylation in mitochondria, chloroplasts and bacteria: natural fuel cells and solar cells. Fed Proc. 1967 Sep;26(5):1370–1379. [PubMed] [Google Scholar]
  17. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  18. Sanderson K. E. Current linkage map of Salmonella typhimurium. Bacteriol Rev. 1970 Jun;34(2):176–193. doi: 10.1128/br.34.2.176-193.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Simoni R. D., Shallenberger M. K. Coupling of energy to active transport of amino acids in Escherichia coli. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2663–2667. doi: 10.1073/pnas.69.9.2663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith H. O., Levine M. A phage P22 gene controlling integration of prophage. Virology. 1967 Feb;31(2):207–216. doi: 10.1016/0042-6822(67)90164-x. [DOI] [PubMed] [Google Scholar]
  21. 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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