Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Feb;129(2):1034–1044. doi: 10.1128/jb.129.2.1034-1044.1977

Mutants of Escherichia coli "cryptic" for certain periplasmic enzymes: evidence for an alteration of the outer membrane.

I R Beacham, D Haas, E Yagil
PMCID: PMC235043  PMID: 320175

Abstract

Mutants in which the expression of periplasmic enzymes by whole cells is reduced (termed "cryptic") are also found to show greatly reduced uptake of labeled adenosine 5'-monophosphate (5'-AMP), providing a rapid assay for crypticity. The crypticity of 3'- and 5'-nucleotidase has been examined as a function of substrate concentration. The Km for 3'- or 5'-AMP increases in the cryptic mutants when whole cells are used as the enzyme source. The Vmax is not altered. Electrophoretic analysis of protein prepared from cell envelopes showed that three cryptic mutants have a polypeptide absent from the outer membrane and a relatively high proportion of a polypeptide in the inner membrane. Analysis of the molar ratios of constituent sugars of the lipopolysaccharides showed no differences between three cryptic mutants and the parent strain. One cryptic mutant (3--41), however, has altered sensitivity to phage T4. By selection for phage resistance, derivatives of the cryptic mutants that are deoxycholate sensitive have been obtained. These mutants are no longer cryptic. We suggest that cryptic mutants have an altered outer membrane, with decreased permeability to 3'- and 5'-AMP, as a result of an altered polypeptide.

Full text

PDF
1034

Images in this article

1039Image
on p.1039
1040Image
on p.1040

Selected References

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

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. ANRAKU Y. A NEW CYCLIC PHOSPHODIESTERASE HAVING A 3'-NUCLEOTIDASE ACTIVITY FROM ESCHERICHIA COLI B. I. PURIFICATION AND SOME PROPERTIES OF THE ENZYME. J Biol Chem. 1964 Oct;239:3412–3419. [PubMed] [Google Scholar]
  3. ARRIGONI O., SINGER T. P. Limitations of the phenazine methosulphate assay for succinic and related dehydrogenases. Nature. 1962 Mar 31;193:1256–1258. doi: 10.1038/1931256a0. [DOI] [PubMed] [Google Scholar]
  4. Ames G. F., Spudich E. N., Nikaido H. Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. J Bacteriol. 1974 Feb;117(2):406–416. doi: 10.1128/jb.117.2.406-416.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beacham I. R., Kahana R., Levy L., Yagil E. Mutants of Escherichia coli K-12 "cryptic," or deficient in 5'-nucleotidase (uridine diphosphate-sugar hydrolase) and 3'-nucleotidase (cyclic phosphodiesterase) activity. J Bacteriol. 1973 Nov;116(2):957–964. doi: 10.1128/jb.116.2.957-964.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Beacham I. R., Silbert D. F. Studies on the uridine diphosphate-galactose: lipopolysaccharide galactosyltransferase reaction using a fatty acid mutant of Escherichia coli. J Biol Chem. 1973 Aug 10;248(15):5310–5318. [PubMed] [Google Scholar]
  8. Boman H. G., Nordström K., Normark S. Penicillin resistance in Escherichia coli K12: synergism between penicillinases and a barrier in the outer part of the envelope. Ann N Y Acad Sci. 1974 May 10;235(0):569–586. doi: 10.1111/j.1749-6632.1974.tb43291.x. [DOI] [PubMed] [Google Scholar]
  9. Bragg P. D., Hou C. Organization of proteins in the native and reformed outer membrane of Escherichia coli. Biochim Biophys Acta. 1972 Aug 9;274(2):478–488. doi: 10.1016/0005-2736(72)90193-9. [DOI] [PubMed] [Google Scholar]
  10. Braun V., Schaller K., Wolff H. A common receptor protein for phage T5 and colicin M in the outer membrane of Escherichia coli B. Biochim Biophys Acta. 1973 Sep 27;323(1):87–97. doi: 10.1016/0005-2736(73)90433-1. [DOI] [PubMed] [Google Scholar]
  11. Brockman R. W., Heppel L. A. On the localization of alkaline phosphatase and cyclic phosphodiesterase in Escherichia coli. Biochemistry. 1968 Jul;7(7):2554–2562. doi: 10.1021/bi00847a016. [DOI] [PubMed] [Google Scholar]
  12. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  13. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  14. Hartree E. F. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972 Aug;48(2):422–427. doi: 10.1016/0003-2697(72)90094-2. [DOI] [PubMed] [Google Scholar]
  15. Henning U., Haller I. Mutants of Escherichia coli K12 lacking all 'major' proteins of the outer cell envelope membrane. FEBS Lett. 1975 Jul 15;55(1):161–164. doi: 10.1016/0014-5793(75)80983-5. [DOI] [PubMed] [Google Scholar]
  16. Koplow J., Goldfine H. Alterations in the outer membrane of the cell envelope of heptose-deficient mutants of Escherichia coli. J Bacteriol. 1974 Feb;117(2):527–543. doi: 10.1128/jb.117.2.527-543.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Leive L. The barrier function of the gram-negative envelope. Ann N Y Acad Sci. 1974 May 10;235(0):109–129. doi: 10.1111/j.1749-6632.1974.tb43261.x. [DOI] [PubMed] [Google Scholar]
  19. Lindberg A. A., Svensson S. Salmonella typhimurium mutations conferring resistance to Felix O phage without loss of smooth character: phage attachment and immunochemical and structural analyses of lipopolysaccharides. J Gen Microbiol. 1975 Mar;87(1):11–19. doi: 10.1099/00221287-87-1-11. [DOI] [PubMed] [Google Scholar]
  20. Lindsay S. S., Wheeler B., Sanderson K. E., Costerton J. W., Cheng K. J. The release of alkaline phosphatase and of lipopolysaccharide during the growth of rough and smooth strains of Salmonella typhimurium. Can J Microbiol. 1973 Mar;19(3):335–343. doi: 10.1139/m73-056. [DOI] [PubMed] [Google Scholar]
  21. MacPhee D. G., Krishnapillai V., Roantree R. J., Stocker B. A. Mutations in Salmonella typhimurium conferring resistance to Felix O phage without loss of smooth character. J Gen Microbiol. 1975 Mar;87(1):1–10. doi: 10.1099/00221287-87-1-1. [DOI] [PubMed] [Google Scholar]
  22. Nakae T. Identification of the outer membrane protein of E. coli that produces transmembrane channels in reconstituted vesicle membranes. Biochem Biophys Res Commun. 1976 Aug 9;71(3):877–884. doi: 10.1016/0006-291x(76)90913-x. [DOI] [PubMed] [Google Scholar]
  23. Nakae T., Nikaido H. Outer membrane as a diffusion barrier in Salmonella typhimurium. Penetration of oligo- and polysaccharides into isolated outer membrane vesicles and cells with degraded peptidoglycan layer. J Biol Chem. 1975 Sep 25;250(18):7359–7365. [PubMed] [Google Scholar]
  24. Nakae T. Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. J Biol Chem. 1976 Apr 10;251(7):2176–2178. [PubMed] [Google Scholar]
  25. Neu H. C. The 5'-nucleotidase of Escherichia coli. I. Purification and properties. J Biol Chem. 1967 Sep 10;242(17):3896–3904. [PubMed] [Google Scholar]
  26. Nikaido H. Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta. 1976 Apr 16;433(1):118–132. doi: 10.1016/0005-2736(76)90182-6. [DOI] [PubMed] [Google Scholar]
  27. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
  30. Schlecht S., Westphal O. Untersuchungen zur Typisierung von Salmonella-R-Formen. 4. Typisierung von S. minnesota-R-Mutanten mittels Antibiotica. Zentralbl Bakteriol Orig. 1970 Apr;213(3):356–380. [PubMed] [Google Scholar]
  31. Schmidt G., Jann B., Jann K. Immunochemistry of R lipopolysaccharides of Escherichia coli. Different core regions in the lipopolysaccharides of O group 8. Eur J Biochem. 1969 Oct;10(3):501–510. doi: 10.1111/j.1432-1033.1969.tb00717.x. [DOI] [PubMed] [Google Scholar]
  32. Schmidt G., Schlecht S., Westphal O. Untersuchungen zur Typisierung von Salmonella-R-Formen. 3. Typisierung von S. minnesota-Mutanten mittels chemischer Agenzien. Zentralbl Bakteriol Orig. 1969 Dec;212(1):88–96. [PubMed] [Google Scholar]
  33. Schnaitman C. A. Outer membrane proteins of Escherichia coli. 3. Evidence that the major protein of Escherichia coli O111 outer membrane consists of four distinct polypeptide species. J Bacteriol. 1974 May;118(2):442–453. doi: 10.1128/jb.118.2.442-453.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schnaitman C. A. Outer membrane proteins of Escherichia coli. I. Effect of preparative conditions on the migration of protein in polyacrylamide gels. Arch Biochem Biophys. 1973 Aug;157(2):541–552. doi: 10.1016/0003-9861(73)90673-5. [DOI] [PubMed] [Google Scholar]
  35. Schnaitman C. A. Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J Bacteriol. 1970 Nov;104(2):890–901. doi: 10.1128/jb.104.2.890-901.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schnaitman C., Smith D., de Salsas M. F. Temperate Bacteriophage Which Causes the Production of a New Major Outer Membrane Protein by Escherichia coli. J Virol. 1975 May;15(5):1121–1130. doi: 10.1128/jvi.15.5.1121-1130.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tamaki S., Sato T., Matsuhashi M. Role of lipopolysaccharides in antibiotic resistance and bacteriophage adsorption of Escherichia coli K-12. J Bacteriol. 1971 Mar;105(3):968–975. doi: 10.1128/jb.105.3.968-975.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ward J. B., Glaser L. An E. coli mutant with cryptic UDP-sugar hydrolase and altered metabolite regulation. Biochem Biophys Res Commun. 1968 Jun 10;31(5):671–677. doi: 10.1016/0006-291x(68)90614-1. [DOI] [PubMed] [Google Scholar]
  39. Ward J. B., Glaser L. Turnover of UDP-sugars in E. Arch Biochem Biophys. 1969 Nov;134(2):612–622. doi: 10.1016/0003-9861(69)90325-7. [DOI] [PubMed] [Google Scholar]
  40. Watson G., Paigen K. Isolation and characterization of an Escherichia coli bacteriophage requiring cell wall galactose. J Virol. 1971 Nov;8(5):669–674. doi: 10.1128/jvi.8.5.669-674.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilkinson R. G., Gemski P., Jr, Stocker B. A. Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J Gen Microbiol. 1972 May;70(3):527–554. doi: 10.1099/00221287-70-3-527. [DOI] [PubMed] [Google Scholar]
  42. Wu H. C. Isolation and characterization of an Escherichia coli mutant with alteration in the outer membrane porteins of the cell envelope. Biochim Biophys Acta. 1972 Dec 1;290(1):274–289. doi: 10.1016/0005-2736(72)90070-3. [DOI] [PubMed] [Google Scholar]
  43. Yagil E., Beacham I. R. Uptake of adenosine 5'-monophosphate by Escherichia coli. J Bacteriol. 1975 Feb;121(2):401–405. doi: 10.1128/jb.121.2.401-405.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES