Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 Oct;144(1):192–199. doi: 10.1128/jb.144.1.192-199.1980

Cation coupling to melibiose transport in Salmonella typhimurium.

S Niiya, Y Moriyama, M Futai, T Tsuchiya
PMCID: PMC294618  PMID: 6998948

Abstract

Melibiose transport in Salmonella typhimurium was investigated. Radioactive melibiose was prepared and the melibiose transport system was characterized. Na+ and Li+ stimulated transport of melibiose by lowering the Km value without affecting the Vmax value; Km values were 0.50 mM in the absence of Na+ or Li+ and 0.12 mM in the presence of 10 mM NaCl or 10 mM LiCl. The Vmax value was 140 nmol/min per mg of protein. Melibiose was a much more effective substrate than methyl-beta-thiogalactoside. An Na+-melibiose cotransport mechanism was suggested by three types of experiments. First, the influx of Na+ induced by melibiose influx was observed with melibiose-induced cells. Second, the efflux of H+ induced by melibiose influx was observed only in the presence of Na+ or Li+, demonstrating the absence of H+-melibiose cotransport. Third, either an artificially imposed Na+ gradient or membrane potential could drive melibiose uptake in cells. Formation of an Na+ gradient in S. typhimurium was shown to be coupled to H+ by three methods. First, uncoupler-sensitive extrusion of Na+ was energized by respiration or glycolysis. Second, efflux of H+ induced by Na+ influx was detected. Third, a change in the pH gradient was elicited by imposing an Na+ gradient in energized membrane vesicles. Thus, it is concluded that the mechanism for Na+ extrusion is an Na+/H+ antiport. The Na+/H+ antiporter is a transformer which converts an electrochemical H+ gradient to an Na+ gradient, which then drives melibiose transport. Li+ was inhibitory for the growth of cells when melibiose was the sole carbon source, even though Li+ stimulated melibiose transport. This suggests that high intracellular Li+ may be harmful.

Full text

PDF

Selected References

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

  1. Beck J. C., Rosen B. P. Cation/proton antiport systems in escherichia coli: properties of the sodium/proton antiporter. Arch Biochem Biophys. 1979 Apr 15;194(1):208–214. doi: 10.1016/0003-9861(79)90611-8. [DOI] [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. Crane R. K. Na+ -dependent transport in the intestine and other animal tissues. Fed Proc. 1965 Sep-Oct;24(5):1000–1006. [PubMed] [Google Scholar]
  4. 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]
  5. Lopilato J., Tsuchiya T., Wilson T. H. Role of Na+ and Li+ in thiomethylgalactoside transport by the melibiose transport system of Escherichia coli. J Bacteriol. 1978 Apr;134(1):147–156. doi: 10.1128/jb.134.1.147-156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. MacDonald R. E., Greene R. V., Lanyi J. K. Light-activated amino acid transport systems in Halobacterium halobium envelope vesicles: role of chemical and electrical gradients. Biochemistry. 1977 Jul 12;16(14):3227–3235. doi: 10.1021/bi00633a029. [DOI] [PubMed] [Google Scholar]
  7. MacDonald R. E., Lanyi J. K., Greene R. V. Sodium-stimulated glutamate uptake in membrane vesicles of Escherichia coli: the role of ion gradients. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3167–3170. doi: 10.1073/pnas.74.8.3167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Saier M. H., Jr, Straud H., Massman L. S., Judice J. J., Newman M. J., Feucht B. U. Permease-specific mutations in Salmonella typhimurium and Escherichia coli that release the glycerol, maltose, melibiose, and lactose transport systems from regulation by the phosphoenolpyruvate:sugar phosphotransferase system. J Bacteriol. 1978 Mar;133(3):1358–1367. doi: 10.1128/jb.133.3.1358-1367.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Schuldiner S., Fishkes H. Sodium-proton antiport in isolated membrane vesicles of Escherichia coli. Biochemistry. 1978 Feb 21;17(4):706–711. doi: 10.1021/bi00597a023. [DOI] [PubMed] [Google Scholar]
  11. Sorensen E. N., Rosen B. P. Effects of sodium and lithium ions on the potassium ion transport systems of Escherichia coli. Biochemistry. 1980 Apr 1;19(7):1458–1462. doi: 10.1021/bi00548a030. [DOI] [PubMed] [Google Scholar]
  12. Stock J., Roseman S. A sodium-dependent sugar co-transport system in bacteria. Biochem Biophys Res Commun. 1971 Jul 2;44(1):132–138. doi: 10.1016/s0006-291x(71)80168-7. [DOI] [PubMed] [Google Scholar]
  13. Tanaka K., Niiya S., Tsuchiya T. Melibiose transport of Escherichia coli. J Bacteriol. 1980 Mar;141(3):1031–1036. doi: 10.1128/jb.141.3.1031-1036.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tanaka S., Lerner S. A., Lin E. C. Replacement of a phosphoenolpyruvate-dependent phosphotransferase by a nicotinamide adenine dinucleotide-linked dehydrogenase for the utilization of mannitol. J Bacteriol. 1967 Feb;93(2):642–648. doi: 10.1128/jb.93.2.642-648.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tokuda H., Kaback H. R. Sodium-dependent methyl 1-thio-beta-D-galactopyranoside transport in membrane vesicles isolated from Salmonella typhimurium. Biochemistry. 1977 May 17;16(10):2130–2136. doi: 10.1021/bi00629a013. [DOI] [PubMed] [Google Scholar]
  16. Tsuchiya T., Hasan S. M., Raven J. Glutamate transport driven by an electrochemical gradient of sodium ions in Escherichia coli. J Bacteriol. 1977 Sep;131(3):848–853. doi: 10.1128/jb.131.3.848-853.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tsuchiya T., Lopilato J., Wilson T. H. Effect of lithium ion on melibiose transport in Escherichia coli. J Membr Biol. 1978 Jul 21;42(1):45–59. doi: 10.1007/BF01870393. [DOI] [PubMed] [Google Scholar]
  18. Tsuchiya T., Raven J., Wilson T. H. Co-transport of Na+ and methul-beta-D-thiogalactopyranoside mediated by the melibiose transport system of Escherichia coli. Biochem Biophys Res Commun. 1977 May 9;76(1):26–31. doi: 10.1016/0006-291x(77)91663-1. [DOI] [PubMed] [Google Scholar]
  19. Tsuchiya T., Takeda K. Calcium/proton and sodium/proton antiport systems in Escherichia coli. J Biochem. 1979 Apr;85(4):943–951. doi: 10.1093/oxfordjournals.jbchem.a132426. [DOI] [PubMed] [Google Scholar]
  20. Tsuchiya T., Takeda K. Extrusion of sodium ions energized by respiration and glycolysis in Escherichia coli. J Biochem. 1979 Jul;86(1):225–230. [PubMed] [Google Scholar]
  21. Tsuchiya T., Takeda K., Wilson T. H. H + -substrate cotransport by the melibiose membrane carrier in Escherichia coli. Membr Biochem. 1980;3(1-2):131–146. doi: 10.3109/09687688009063881. [DOI] [PubMed] [Google Scholar]
  22. Tsuchiya T., Wilson T. H. Cation-sugar cotransport in the melibiose transport system of Escherichia coli. Membr Biochem. 1978;2(1):63–79. doi: 10.3109/09687687809063858. [DOI] [PubMed] [Google Scholar]
  23. West I. C., Mitchell P. Proton/sodium ion antiport in Escherichia coli. Biochem J. 1974 Oct;144(1):87–90. doi: 10.1042/bj1440087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wilson T. H., Maloney P. C. Speculations on the evolution of ion transport mechanisms. Fed Proc. 1976 Aug;35(10):2174–2179. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES