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.
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)
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)
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)
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)
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)
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)
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)
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)
Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K12. Bacteriol. Rev. 40, 116–167 (1976)
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)
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)
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)
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)
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)
Boman, H.G., Monner, D.A.: Characterization of lipopolysaccharides from Escherichia coli K-12 mutants. J. Bacteriol. 121, 455–464 (1975)
Braun, V.: Covalent lipoprotein from the outer membrane of E. coli. Biochim. Biophys. Acta (Amst.) 415, 335–377 (1975)
Chai, T., Foulds, J.: Escherichia coli K12 tolF mutants: alterations in protein composition of the outer membrane. J. Bacteriol. 130, 781–786 (1977)
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)
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)
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)
Foulds, J.: tolF locus in Escherichia coli: chromosomal location and relationship to loci cmlB and tolD. J. Bacteriol. 128, 604–608 (1976)
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)
Galanos, C., Lüderitz, O., Westphal, O.: A new method for the extraction of R lipopolysaccharides. Eur. J. Biochem. 9, 245–249 (1969)
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)
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
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)
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)
Henning, U., Höhn, B., Sonntag, I.: Cell envelope and shape of Escherichia coli K12. The ghost membrane. Eur. J. Biochem. 39, 27–36 (1973)
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)
Hindennach, I., Henning, U.: The major proteins of the Escherichia coli outer cell envelope membrane. Eur. J. Biochem. 59, 207–213 (1975)
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)
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)
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)
Lugtenberg, B., Bronstein, H., van Selm, N., Peters, R.: Peptidoglycanassociated outer membrane proteins in Gramnegative bacteria. Biochim. Biophys. Acta (Amst.) 465, 571–578 (1977)
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)
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)
Lutkenhaus, J.F.: Role of major outer membrane protein in Escherichia coli. J. Bacteriol. 131, 631–637 (1977)
Meyenburg, K. von: Transport-limited growth rates in a mutant of Escherichia coli. J. Bacteriol. 107, 878–888 (1971)
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)
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)
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)
Sarma, V., Reeves, P.: Genetic locus (ompB) affecting a major outer-membrane protein in Escherichia coli K12. J. Bacteriol. 132, 23–27 (1977)
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)
Schnaitman, C.A.: Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J. Bacteriol. 104, 890–901 (1970)
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)
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)
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)
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)
Tarr, G.E.: A general procedure for the manual sequencing of small quantities of peptides. Anal. Biochem. 63, 361–370 (1975)
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)
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)
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)
Author information
Authors and Affiliations
Additional information
Communicated by H.G. Wittmann
Rights 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
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00271664