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. 1983 Jul;155(1):186–195. doi: 10.1128/jb.155.1.186-195.1983

sn-Glycerol-3-phosphate transport in Salmonella typhimurium.

R Hengge, T J Larson, W Boos
PMCID: PMC217668  PMID: 6408060

Abstract

Salmonella typhimurium contains a transport system for sn-glycerol-3-phosphate that is inducible by growth on glycerol and sn-glycerol-3-phosphate. In fully induced cells, the system exhibited an apparent Km of 50 microM and a Vmax of 2.2 nmol/min . 10(8) cells. The corresponding system in Escherichia coli exhibits, under comparable conditions, a Km of 14 microM and a Vmax of 2.2 nmol/min . 10(8) cells. Transport-defective mutants were isolated by selecting for resistance against the antibiotic fosfomycin. They mapped in glpT at 47 min in the S. typhimurium linkage map, 37% cotransducible with gyrA. In addition to the glpT-dependent system, S. typhimurium LT2 contains, like E. coli, a second, ugp-dependent transport system for sn-glycerol-3-phosphate that was derepressed by phosphate starvation. A S. typhimurium DNA bank containing EcoRI restriction fragments in phage lambda gt7 was used to clone the glpT gene in E. coli. Lysogens that were fully active in the transport of sn-glycerol-3-phosphate with a Km of 33 microM and a Vmax of 2.0 nmol/min . 10(8) cells were isolated in a delta glpT mutant of E. coli. The EcoRI fragment harboring glpT was 3.5 kilobases long and carried only part of glpQ, a gene distal to glpT but on the same operon. The fragment was subcloned in multicopy plasmid pACYC184. Strains carrying this hybrid plasmid produced large amounts of cytoplasmic membrane protein with an apparent molecular weight of 33,000, which was identified as the sn-glycerol-3-phosphate permease. Its properties were similar to the corresponding E. coli permease. The presence of the multicopy glpT hybrid plasmid had a strong influence on the synthesis or assembly of other cell envelope proteins of E. coli. For instance, the periplasmic ribose-binding protein was nearly absent. On the other hand, the quantity of an unidentified E. coli outer membrane protein usually present only in small amounts increased.

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Selected References

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

  1. Aksamit R., Koshland D. E., Jr A ribose binding protein of Salmonella typhimurium. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1348–1353. doi: 10.1016/0006-291x(72)90860-1. [DOI] [PubMed] [Google Scholar]
  2. Alper M. D., Ames B. N. Cyclic 3', 5'-adenosine monophosphate phosphodiesterase mutants of Salmonella typhimurium. J Bacteriol. 1975 Jun;122(3):1081–1090. doi: 10.1128/jb.122.3.1081-1090.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alper M. D., Ames B. N. Transport of antibiotics and metabolite analogs by systems under cyclic AMP control: positive selection of Salmonella typhimurium cya and crp mutants. J Bacteriol. 1978 Jan;133(1):149–157. doi: 10.1128/jb.133.1.149-157.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Argast M., Boos W. Co-regulation in Escherichia coli of a novel transport system for sn-glycerol-3-phosphate and outer membrane protein Ic (e, E) with alkaline phosphatase and phosphate-binding protein. J Bacteriol. 1980 Jul;143(1):142–150. doi: 10.1128/jb.143.1.142-150.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Argast M., Schumacher G., Boos W. Characterization of a periplasmic protein related to sn-glycerol-3-phosphate transport in escherichia coli. J Supramol Struct. 1977;6(1):135–153. doi: 10.1002/jss.400060111. [DOI] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Boos W., Hartig-Beecken I., Altendorf K. Purification and properties of a periplasmic protein related to sn-glycerol-3-phosphate transport in Escherichia coli. Eur J Biochem. 1977 Feb;72(3):571–581. doi: 10.1111/j.1432-1033.1977.tb11280.x. [DOI] [PubMed] [Google Scholar]
  8. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cordaro J. C., Melton T., Stratis J. P., Atagün M., Gladding C., Hartman P. E., Roseman S. Fosfomycin resistance: selection method for internal and extended deletions of the phosphoenolpyruvate:sugar phosphotransferase genes of Salmonella typhimurium. J Bacteriol. 1976 Dec;128(3):785–793. doi: 10.1128/jb.128.3.785-793.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Emr S. D., Hanley-Way S., Silhavy T. J. Suppressor mutations that restore export of a protein with a defective signal sequence. Cell. 1981 Jan;23(1):79–88. doi: 10.1016/0092-8674(81)90272-5. [DOI] [PubMed] [Google Scholar]
  11. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  12. Hendlin D., Stapley E. O., Jackson M., Wallick H., Miller A. K., Wolf F. J., Miller T. W., Chaiet L., Kahan F. M., Foltz E. L. Phosphonomycin, a new antibiotic produced by strains of streptomyces. Science. 1969 Oct 3;166(3901):122–123. doi: 10.1126/science.166.3901.122. [DOI] [PubMed] [Google Scholar]
  13. Herrero E., Jackson M., Bassford P. J., Sinden D., Holland I. B. Insertion of a MalE beta-galactosidase fusion protein into the envelope of Escherichia coli disrupts biogenesis of outer membrane proteins and processing of inner membrane proteins. J Bacteriol. 1982 Oct;152(1):133–139. doi: 10.1128/jb.152.1.133-139.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ito K., Bassford P. J., Jr, Beckwith J. Protein localization in E. coli: is there a common step in the secretion of periplasmic and outer-membrane proteins? Cell. 1981 Jun;24(3):707–717. doi: 10.1016/0092-8674(81)90097-0. [DOI] [PubMed] [Google Scholar]
  15. Johnson W. C., Silhavy T. J., Boos W. Two-dimensional polyacylamide gel electrophoresis of envelope proteins of Escherichia coli. Appl Microbiol. 1975 Mar;29(3):405–413. doi: 10.1128/am.29.3.405-413.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kahan F. M., Kahan J. S., Cassidy P. J., Kropp H. The mechanism of action of fosfomycin (phosphonomycin). Ann N Y Acad Sci. 1974 May 10;235(0):364–386. doi: 10.1111/j.1749-6632.1974.tb43277.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Larson T. J., Ehrmann M., Boos W. Periplasmic glycerophosphodiester phosphodiesterase of Escherichia coli, a new enzyme of the glp regulon. J Biol Chem. 1983 May 10;258(9):5428–5432. [PubMed] [Google Scholar]
  20. Larson T. J., Schumacher G., Boos W. Identification of the glpT-encoded sn-glycerol-3-phosphate permease of Escherichia coli, an oligomeric integral membrane protein. J Bacteriol. 1982 Dec;152(3):1008–1021. doi: 10.1128/jb.152.3.1008-1021.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ludtke D., Larson T. J., Beck C., Boos W. Only one gene is required for the glpT-dependent transport of sn-glycerol-3-phosphate in Escherichia coli. Mol Gen Genet. 1982;186(4):540–547. doi: 10.1007/BF00337962. [DOI] [PubMed] [Google Scholar]
  22. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  23. Richarme G., Kepes A. Release of glucose from purified galactose-binding protein of Escherichia coli upon addition of galactose. Eur J Biochem. 1974 Jun 1;45(1):127–133. doi: 10.1111/j.1432-1033.1974.tb03537.x. [DOI] [PubMed] [Google Scholar]
  24. Schryvers A., Weiner J. H. The anaerobic sn-glycerol-3-phosphate dehydrogenase: cloning and expression of the glpA gene of Escherichia coli and identification of the glpA products. Can J Biochem. 1982 Mar;60(3):224–231. doi: 10.1139/o82-027. [DOI] [PubMed] [Google Scholar]
  25. Schweizer H., Argast M., Boos W. Characteristics of a binding protein-dependent transport system for sn-glycerol-3-phosphate in Escherichia coli that is part of the pho regulon. J Bacteriol. 1982 Jun;150(3):1154–1163. doi: 10.1128/jb.150.3.1154-1163.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schweizer H., Grussenmeyer T., Boos W. Mapping of two ugp genes coding for the pho regulon-dependent sn-glycerol-3-phosphate transport system of Escherichia coli. J Bacteriol. 1982 Jun;150(3):1164–1171. doi: 10.1128/jb.150.3.1164-1171.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shultz J., Silhavy T. J., Berman M. L., Fiil N., Emr S. D. A previously unidentified gene in the spc operon of Escherichia coli K12 specifies a component of the protein export machinery. Cell. 1982 Nov;31(1):227–235. doi: 10.1016/0092-8674(82)90422-6. [DOI] [PubMed] [Google Scholar]
  28. Silhavy T. J., Hartig-Beecken I., Boos W. Periplasmic protein related to the sn-glycerol-3-phosphate transport system of Escherichia coli. J Bacteriol. 1976 May;126(2):951–958. doi: 10.1128/jb.126.2.951-958.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Venkateswaran P. S., Wu H. C. Isolation and characterization of a phosphonomycin-resistant mutant of Escherichia coli K-12. J Bacteriol. 1972 Jun;110(3):935–944. doi: 10.1128/jb.110.3.935-944.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. West I., Mitchell P. Proton-coupled beta-galactoside translocation in non-metabolizing Escherichia coli. J Bioenerg. 1972 Aug;3(5):445–462. doi: 10.1007/BF01516082. [DOI] [PubMed] [Google Scholar]
  31. Winkler H. H. Distribution of an inducible hexose-phosphate transport system among various bacteria. J Bacteriol. 1973 Nov;116(2):1079–1081. doi: 10.1128/jb.116.2.1079-1081.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yagil E., Hermoni E. Repression of alkaline phosphatase in Salmonella typhimurium carrying a phoA+ phoR- episome from Escherichia coli. J Bacteriol. 1976 Nov;128(2):661–664. doi: 10.1128/jb.128.2.661-664.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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