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. 1975 Sep;123(3):928–936. doi: 10.1128/jb.123.3.928-936.1975

Membrane-associated nucleotide sugar reactions: influence of mutations affecting lipopolysaccharide on the first enzyme of O-antigen synthesis.

K Rundell, C W Shuster
PMCID: PMC235816  PMID: 1099085

Abstract

Both the synthesis of lipopolysaccharide O-antigen and the synthesis of peptidoglycan in Salmonella typhimurium proceed via membrane-bound glycosylated lipid intermediates. The first enzyme of each pathway transfers a sugar phosphate from a nucleotide sugar to the glycosyl carrier lipid (P-GCL). Each enzyme catalyzes an exchange reaction between the reaction product urine monophosphate, and the nucleotide sugar substrate. Several strains of S. typhimurium defective in lipopolysaccharide synthesis accumulate glycosylated lipid intermediates under appropriate conditions. In addition, strains lysogenic for phage P22 synthesize a glucose derivative of the carrier lipid. These strains were used to demonstrate the P/GCL requirement of the exchange reaction catalyzed by galactose-diphosphoglycosyl carrier lipid (GCL-PP-Gal) synthetase, the first enzyme of O-antigen synthesis. Enzyme activity is greatly reduced when glycosylated P-GCL accumulates on the cytoplasmic membrane. The exchange reaction catalyzed by the first enzyme of peptidoglycan synthesis is unaffected by the accumulation of O-antigen fragments on the carrier lipid and may interact with a different pool of P-GCL within the membrane. GCL-PP-Gal synthetase activity cannot be detected in the membranes of two rfa mutants that synthesize incomplete lipopolysaccharide core. Either the synthesis of GCL-PP-Gal synthetase or the stable integration of the enzyme into the membrane structure may be disrupted in the rfa mutants. Peptidoglycan synthesis is unaffected by the mutations affecting the core glycosyltransferases.

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Selected References

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

  1. Ames G. F. Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism. J Bacteriol. 1968 Mar;95(3):833–843. doi: 10.1128/jb.95.3.833-843.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dankert M., Wright A., Kelley W. S., Robbins P. W. Isolation, purification, and properties of the lipid-linked intermediates of O-antigen biosynthesis. Arch Biochem Biophys. 1966 Sep 26;116(1):425–435. doi: 10.1016/0003-9861(66)90049-x. [DOI] [PubMed] [Google Scholar]
  3. FUKAZAWA Y., HARTMAN P. E. A P22 BACTERIOPHAGE MUTANT DEFECTIVE IN ANTIGEN CONVERSION. Virology. 1964 Jun;23:279–283. doi: 10.1016/0042-6822(64)90296-x. [DOI] [PubMed] [Google Scholar]
  4. Higashi Y., Strominger J. L., Sweeley C. C. Biosynthesis of the peptidoglycan of bacterial cell walls. XXI. Isolation of free C55-isoprenoid alcohol and of lipid intermediates in peptidoglycan synthesis from Staphylococcus aureus. J Biol Chem. 1970 Jul 25;245(14):3697–3702. [PubMed] [Google Scholar]
  5. Hinckley, Müller E., Rothfield L. Reassembly of a membrane-bound multienzyme system. I. Formation of a particle containing phosphatidylethanolamine, lipopolysaccharide, and two glycosyltransferase enzymes. J Biol Chem. 1972 Apr 25;247(8):2623–2628. [PubMed] [Google Scholar]
  6. Kuo T. T., Stocker B. A. Mapping of rfa Genes in Salmonella typhimurium by ES18 and P22 Transduction and by Conjugation. J Bacteriol. 1972 Oct;112(1):48–57. doi: 10.1128/jb.112.1.48-57.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Nikaido H. Biosynthesis of cell wall lipopolysaccharide in gram-negative enteric bacteria. Adv Enzymol Relat Areas Mol Biol. 1968;31:77–124. doi: 10.1002/9780470122761.ch3. [DOI] [PubMed] [Google Scholar]
  9. Nikaido H., Nikaido K., Nakae T., Mäkelä P. H. Glucosylation of lipopolysaccharide in Salmonella: biosynthesis of O antigen factor 12 2 . I. Over-all reaction. J Biol Chem. 1971 Jun 25;246(12):3902–3911. [PubMed] [Google Scholar]
  10. Osborn M. J. Biochemical characterization of mutants of Salmonella typhimurium lacking glucosyl or galactosyl lipopolysaccharide transferases. Nature. 1968 Mar 9;217(5132):957–960. doi: 10.1038/217957a0. [DOI] [PubMed] [Google Scholar]
  11. Osborn M. J., Weiner I. M. Biosynthesis of a bacterial lipopolysaccharide. VI. Mechanism of incorporation of abequose into the O-antigen of Salmonella typhimurium. J Biol Chem. 1968 May 25;243(10):2631–2639. [PubMed] [Google Scholar]
  12. Pless D. D., Neuhaus F. C. Initial membrane reaction in peptidoglycan synthesis. Lipid dependence of phospho-n-acetylmuramyl-pentapeptide translocase (exchange reaction). J Biol Chem. 1973 Mar 10;248(5):1568–1576. [PubMed] [Google Scholar]
  13. Rundell K., Shuster C. W. Membrane-associated nucleotide sugar reactions. I. Properties of the first enzyme of O antigen synthesis. J Biol Chem. 1973 Aug 10;248(15):5436–5442. [PubMed] [Google Scholar]
  14. STOCKER B. A., STAUB A. M., TENELLI R., KOPACKA B. [Immunochemical study on Salmonella. VI. Study of antigen 1 present on 2 Salmonella of the B and E4 groups]. Ann Inst Pasteur (Paris) 1960 Apr;98:505–523. [PubMed] [Google Scholar]
  15. SUBBAIAH T. V., STOCKER B. A. ROUGH MUTANTS OF SALMONELLA TYPHIMURIUM. I. GENETICS. Nature. 1964 Mar 28;201:1298–1299. doi: 10.1038/2011298a0. [DOI] [PubMed] [Google Scholar]
  16. Sanderson K. E. Linkage map of Salmonella typhimurium, edition IV. Bacteriol Rev. 1972 Dec;36(4):558–586. doi: 10.1128/br.36.4.558-586.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shuster C. W., Rundell K. Resistance of Salmonella typhimurium mutants to galactose death. J Bacteriol. 1969 Oct;100(1):103–109. doi: 10.1128/jb.100.1.103-109.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stickgold R. A., Neuhaus F. C. On the initial stage in peptidoglycan synthesis. Effect of 5-fluorouracil substitution on phospho-N-acetylmuramyl-pentapeptide translocase (uridine 5'-phosphate). J Biol Chem. 1967 Mar 25;242(6):1331–1337. [PubMed] [Google Scholar]
  19. Struve W. G., Sinha R. K., Neuhaus F. C. On the initial stage in peptidoglycan synthesis. Phospho-N-acetylmuramyl-pentapeptide translocase (uridine monophosphate). Biochemistry. 1966 Jan;5(1):82–93. doi: 10.1021/bi00865a012. [DOI] [PubMed] [Google Scholar]
  20. Weiner I. M., Higuchi T., Osborn M. J., Horecker B. L. Biosynthesis of O-antigen in Salmonella typhimurium. Ann N Y Acad Sci. 1966 Jun 30;133(2):391–404. doi: 10.1111/j.1749-6632.1966.tb52379.x. [DOI] [PubMed] [Google Scholar]

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