Log in

Genetics and biochemistry of the peptidoglycan-associated proteins b and c of Escherichia coli K12

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

To collect information on synthesis and regulation of the peptidoglycan-associated pore-forming outer membrane proteins b and c, mutants resistant to phages Mel and TuIa were analyzed. Genetic analysis showed three linkage groups, corresponding with the genes tolF (phenotype b-c+), meo A (phenotype b+c-) and ompB (phenotypes b-c-, b- c+, b++ c- and b++ c±). It has recently been described that also a b+ c- phenotype can occur in the latter linkage group [Chai, T., Foulds, J., J. Bacteriol. 130, 781–786 (1977)]. Among ompB (b- c+)/meoA (b+ c-) double mutants strains were found with the b+ c- phenotype, showing that ompB is not the structural gene for protein b. Studies on purified proteins b and c showed profound differences between the two proteins with respect to the electrophoretic mobility of fragments obtained by treatment with cyanogen bromide, trypsin and chymotrypsin. The amino acid in position three of the amino-termini of proteins b and c, isolated from isogenic strains, were identified as isoleucine and valine respectively. Both the genetic and biochemical results are consistent with a model recently published [Ichihara, S., Mizushima, S., J. Biochem. (Japan) 83, 1095–1100 (1978)] which predicts that tolF and meoA are the structural genes for the proteins b and c respectively and that ompB is a regulatory gene whose product regulates the levels of both proteins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alphen, W. van, Boxtel, R. van, Selm, N. van, Lugtenberg, B.: Pores in the outer membrane of Escherichia coli K12. Involvement of proteins b and c in the permeation of cephaloridine and ampicillin. FEMS Lett. 3, 103–106 (1978b)

    Google Scholar 

  • Alphen, W. van, Lugtenberg, B.: Influence of the osmolarity of the growth medium on the outer membrane protein pattern of Escherichia coli K12. J. Bacteriol. 131, 623–630 (1977)

    Google Scholar 

  • Alphen, W. van, Lugtenberg, B., Berendsen, W.: Heptose-deficient mutants of Escherichia coli K12 deficient in up to three major outer membrane proteins. Mol. Gen. Genet. 147, 263–269 (1976)

    Google Scholar 

  • Alphen, L. van, Lugtenberg, B., Boxtel, R. van, Hack, A., Verhoef, C., Havekes, L.: meo A is the structural gene for outer membrane protein c of Escherichia coli K12. Mol. Gen. Genet. 169, 147–155 (1979)

    Google Scholar 

  • Alphen, L. van, Lugtenberg, B., Boxtel, R. van, Verhoef, C.: Architecture of the outer membrane of Escherichia coli K12. I. Action of phospholipases A2 and C on wild type strains and outer membrane mutants. Biochim. Biophys. Acta (Amst.) 466, 257–268 (1977)

    Google Scholar 

  • Alphen, W. van, Selm, N. van, Lugtenberg, B.: Pores in the outer membrane of Escherichia coli K12. Involvement of proteins b and e in the functioning of pores for nucleotides. Mol. Gen. Genet. 159, 75–83 (1978a)

    Google Scholar 

  • Alphen, L. van, Verkleij, A., Leunissen-Bijvelt, J., Lugtenberg, B.: Architecture of the outer membrane of Escherichia coli K12. III. Protein-lipopolysaccharide complexes in intramembraneous particles. J. Bacteriol. 134, 1089–1098 (1978)

    Google Scholar 

  • Ames, G.F., Spudich, E.N., Nikaido, H.: Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. J. Bacteriol. 117, 406–416 (1974)

    Google Scholar 

  • Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K12. Bacteriol. Rev. 40, 116–167 (1976)

    Google Scholar 

  • Bailey, G.S., Gillett, D., Hill, D.F., Petersen, G.B.: Automated sequencing of insoluble peptides using detergent. J. Biol. Chem. 252, 2218–2225 (1977)

    Google Scholar 

  • Bassford, P. J., Jr, Diedrich, D. L., Schnaitman, C.A., Reeves, P.: Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants. J. Bacteriol. 131, 608–622 (1977)

    Google Scholar 

  • Bavoil, P., Nikaido, H., von Meyenburg, K.: Pleiotropic transport mutants of Escherichia coli lack porin, a major outer membrane protein. Mol. Gen. Genet. 158, 23–33 (1978)

    Google Scholar 

  • Beacham, I.R., Haas, O., Yagil, E.: Mutants of Escherichia coli “cryptic” for certain periplasmic enzymes: evidence for an alteration of the outer membrane. J. Bacteriol. 129, 1034–1044 (1977)

    Google Scholar 

  • Beacham, I.R., Kahana, R., Levy, L., Yagil, E.: Mutants of Escherichia coli K12 “cryptic”, or deficient in 5′-nucleotidase (uridine diphosphate sugar hydrolase) and 3′-nucleotidase (cyclic phosphodiesterase) activity. J. Bacteriol. 116, 957–964 (1973)

    Google Scholar 

  • Boman, H.G., Monner, D.A.: Characterization of lipopolysaccharides from Escherichia coli K-12 mutants. J. Bacteriol. 121, 455–464 (1975)

    Google Scholar 

  • Braun, V.: Covalent lipoprotein from the outer membrane of E. coli. Biochim. Biophys. Acta (Amst.) 415, 335–377 (1975)

    Google Scholar 

  • Chai, T., Foulds, J.: Escherichia coli K12 tolF mutants: alterations in protein composition of the outer membrane. J. Bacteriol. 130, 781–786 (1977)

    Google Scholar 

  • Cleveland, D.W., Fischer, S.G., Kirschner, M.W., Laemmli, U.K.: Peptide map** by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J. Biol. Chem. 252, 1102–1106 (1977)

    Google Scholar 

  • Davies, J.K., Reeves, P.: Genetics of resistance to colicins in Escherichia coli K-12: cross resistance among colicins of group A. J. Bacteriol. 123, 102–117 (1975)

    Google Scholar 

  • Dietrich, D.L., Summers, A.O., Schnaitman, C.A.: Outer membrane proteins of Escherichia coli. V. Evidence that protein 1 and bacteriophage-directed protein 2 are different polypeptides. J. Bacteriol. 131, 598–607 (1977)

    Google Scholar 

  • Foulds, J.: tolF locus in Escherichia coli: chromosomal location and relationship to loci cmlB and tolD. J. Bacteriol. 128, 604–608 (1976)

    Google Scholar 

  • Foulds, J., Chai, T.: New major outer membrane protein found in an Escherichia coli tolF mutant resistant to bacteriophage Tulb. J. Bacteriol. 133, 1478–1483 (1978)

    Google Scholar 

  • Galanos, C., Lüderitz, O., Westphal, O.: A new method for the extraction of R lipopolysaccharides. Eur. J. Biochem. 9, 245–249 (1969)

    Google Scholar 

  • Garten, W., Hindennach, I., Henning, U.: The major proteins of the Escherichia coli outer cell-envelope membrane. Cyanogen bromide fragments of protein I, composition and order. Eur. J. Biochem. 60, 303–307 (1975)

    Google Scholar 

  • Gray, W.R.: Sequence analysis with dansyl chloride. In: Methods in enzymology, 25B (Hirs, C.H.W., Timasheff, S.N., eds.), pp. 333–344 New York: Academic Press 1972

    Google Scholar 

  • Havekes, L. M., Lugtenberg, B.J.J., Hoekstra, W.P.M.: Conjugation deficient E. coli K12 F- mutants with heptose-less lipopolysaccharide. Mol. Gen. Genet. 146, 43–50 (1976)

    Google Scholar 

  • Henning, U., Haller, I.: Mutants of Escherichia coli K12 lacking all ‘major’ proteins of the outer cell envelope membrane. FEBS Lett. 55, 161–164 (1975)

    Google Scholar 

  • Henning, U., Höhn, B., Sonntag, I.: Cell envelope and shape of Escherichia coli K12. The ghost membrane. Eur. J. Biochem. 39, 27–36 (1973)

    Google Scholar 

  • Henning, U., Schmidmayr, W., Hindennach, I.: Major proteins of the outer cell envelope membrane of Escherichia coli K12: multiple species of protein I. Mol. Gen. Genet. 154, 293–298 (1977)

    Google Scholar 

  • Hindennach, I., Henning, U.: The major proteins of the Escherichia coli outer cell envelope membrane. Eur. J. Biochem. 59, 207–213 (1975)

    Google Scholar 

  • Ichihara, S., Mizushima, S.: Characterization of major outer membrane proteins O8 and O9 of Escherichia coli K12. Evidence that structural genes for the two proteins are different. J. Biochem. (Tokyo) 83, 1095–1100 (1978)

    Google Scholar 

  • Inouye, M., Shaw, S., Shen, C.: The assembly of a structural lipoprotein in the envelope of Escherichia coli. J. Biol. Chem. 247, 8154–8159 (1972)

    Google Scholar 

  • Koplow, J., Goldfine, H.: Alterations in the outer membrane of the cell envelope of heptose-deficient mutants of Escherichia coli. J. Bacteriol. 117, 527–543 (1974)

    Google Scholar 

  • Lugtenberg, B., Bronstein, H., van Selm, N., Peters, R.: Peptidoglycanassociated outer membrane proteins in Gramnegative bacteria. Biochim. Biophys. Acta (Amst.) 465, 571–578 (1977)

    Google Scholar 

  • Lugtenberg, B., Meijers, J., Peters, R., van der Hoeck, P., van Alphen, L.: Electrophoretic resolution of the ‘major outer membrane protein’ of Escherichia coli K12 into four bands. FEBS-Lett. 58, 254–258 (1975)

    Google Scholar 

  • Lugtenberg, B., Peters, R., Bernheimer, H., Berendsen, W.: Influence of cultural conditions and mutations on the composition of the outer membrane proteins of Escherichia coli. Mol. Gen. Genet. 147, 251–262 (1976)

    Google Scholar 

  • Lutkenhaus, J.F.: Role of major outer membrane protein in Escherichia coli. J. Bacteriol. 131, 631–637 (1977)

    Google Scholar 

  • Meyenburg, K. von: Transport-limited growth rates in a mutant of Escherichia coli. J. Bacteriol. 107, 878–888 (1971)

    Google Scholar 

  • Nakae, T.: Identification of the outer membrane protein of E. coli that produces transmembrane channels in reconstituted vesicle membranes. Biochem. Biophys. Res. Commun. 71, 877–884 (1976)

    Google Scholar 

  • Nakamura, K., Mizushima, S.: Effects of heating in dodecyl sulfate solution on the conformation and electrophoretic mobility of isolated major outer membrane proteins from Escherichia coli K12. J. Biochem. (Tokyo) 80, 1411–1422 (1976)

    Google Scholar 

  • Rosenbusch, J.P.: Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulphate binding. J. Biol. Chem. 249, 8019–8029 (1974)

    Google Scholar 

  • Sarma, V., Reeves, P.: Genetic locus (ompB) affecting a major outer-membrane protein in Escherichia coli K12. J. Bacteriol. 132, 23–27 (1977)

    Google Scholar 

  • Schmitges, C.J., Henning, U.: The major proteins of the Escherichia coli outer cell-envelope membrane. Heterogeneity of protein I. Eur. J. Biochem. 63, 47–52 (1976)

    Google Scholar 

  • Schnaitman, C.A.: Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J. Bacteriol. 104, 890–901 (1970)

    Google Scholar 

  • Schnaitman, C.A.: Outer membrane proteins of Escherichia coli. IV. Differences in outer membrane proteins due to strain and cultural differences. J. Bacteriol. 118, 454–464 (1974)

    Google Scholar 

  • Schnaitman, C.A., Smith, D., Forn de Salsas, M.: Temperate bacteriophage which causes the production of a new major outer membrane protein by Escherichia coli. J. Virol. 15, 1121–1130 (1975)

    Google Scholar 

  • Schroeder, W.A., Shelton, J.B., Shelton, J. R.: An examination of conditions for the cleavage of polypeptide chains with cyanogen bromide: application to catalase. Arch. Biochem. Biophys. 130, 551–556 (1969)

    Google Scholar 

  • Skurray, R.A., Hancock, R.E.W., Reeves, P.: Con mutants: class of mutants in Escherichia coli K12 lacking a major cell wall protein and defective in conjugation and adsorption of a bacteriophage. J. Bacteriol. 119, 726–735 (1974)

    Google Scholar 

  • Tarr, G.E.: A general procedure for the manual sequencing of small quantities of peptides. Anal. Biochem. 63, 361–370 (1975)

    Google Scholar 

  • Uemura, J., Mizushima, S.: Isolation of outer membrane proteins of E. coli and their characterization on polyacrylamide gel. Biochim. Biophys. Acta (Amst.) 413, 163–176 (1975)

    Google Scholar 

  • Verhoef, C., de Graaff, P.J., Lugtenberg, B.: Map** of a gene for a major outer membrane protein of Escherichia coli K12 with the aid of a newly isolated bacteriophage. Mol. Gen. Genet. 150, 103–105 (1977)

    Google Scholar 

  • Yu, F., Mizushima, S.: Stimulation by lipopolysaccharide of the binding of outer membrane proteins 0–8 and 0–9 to the peptidoglycan layer of Escherichia coli K-12. Biochem. Biophys. Res. Commun. 74, 1397–1402 (1977)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H.G. Wittmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Verhoef, C., Lugtenberg, B., van Boxtel, R. et al. Genetics and biochemistry of the peptidoglycan-associated proteins b and c of Escherichia coli K12. Molec. Gen. Genet. 169, 137–146 (1979). https://doi.org/10.1007/BF00271664

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00271664

Keywords

Navigation