Advertisement

Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Pleiotropic transport mutants of Escherichia coli lack porin, a major outer membrane protein

  • 79 Accesses

  • 68 Citations

Summary

Four “pleiotropic transport” mutants of Escherichia coli B/r with decreased affinity for the uptake of most nutrients were found to lack a major outer membrane protein of 36,500 daltons (“porin”) previously shown to produce transmembrane diffusion channels in in vitro reconstitution experiments. Consequent decrease in outer membrane permeability was confirmed by measuring the transmembrane diffusion rate of 6-aminopenicillanic acid. Quantitative considerations on the porin-dependent permeability of the outer membrane show that (a) there may be very large differences in the actual rates of penetration, even among the “permeable” substances and (b) the numbers of porin molecules present in wild type cells is several orders of magnitude higher than that necessary for the uptake of rapidly diffusing substrates such as glocose from ordinary culture media. The absence of porin and the pleiotropic transport defect were always contransduced, and the mutation was mapped at 73.7 min between aroB and malT by P1 transduction. When “revertants” able to grow on low concentrations of lactose were selected, in addition to true revertants “suppressor” strains with increased amounts of non-porin membrane proteins were isolated.

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

References

  1. Ames, G.F.-L., Nikaido, K.: Two dimensional gel electrophoresis of membrane proteins. Biochemistry 15, 616–623 (1976)

  2. Ames, G.F.-L., Spudich, E.N., Nikaido, H.: Protein composition of the outer membrane of Salmonella typhimurium: Effect of lipopolysaccharide mutations. J. Bact. 117, 406–416 (1974)

  3. Anderson, J.J., Oxender, D.L.: Escherichia coli transport mutants lacking binding protein and other components of the branched chain amino acid transport systems. J. Bact. 130, 384–392 (1977)

  4. Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Genetics 76, 169–184 (1976)

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

  6. Bertani, G.: Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bact. 62, 293–300 (1951)

  7. Boyer, H.: Conjugation in E. coli. J. Bact. 91, 1767–1772 (1966)

  8. Decad, G.M., Nikaido, H.: Outer membrane of Gram-negative bacteria. XII. Molecular sieving function of cell wall. J. Bact. 128, 325–336 (1976)

  9. Hancock, R.E.W., Hantke, K., Braun, V.: Iron transport in Escherichia coli K-12: Involvement of the colicin B receptor and of a citrate-inducible protein. J. Bact. 127, 1370–1375 (1976)

  10. Hantke, K.: Phage T6-colicin K receptor and nucleoside transport in Escherichia coli. FEBS Lett. 70, 109–112 (1976)

  11. Hatfield, D., Hofnung, M., Schwartz, M.: Genetic analysis of the maltose A region in Escherichia coli. J. Bact. 98, 559–567 (1969)

  12. 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)

  13. Hindennach, I., Henning, U.: The major proteins of the Escherichia coli outer cell envelope membrane. Preparative isolation of all major proteins. Europ. J. Biochem. 59, 207–213 (1975)

  14. Hofnung, M.: Divergent operons and the genetic structure of the maltose B region of Escherichia coli K12. Genetics 76, 169–184 (1974)

  15. Ichihara, S., Mizushima, S.: Involvement of outer membrane proteins in enterochelin-mediated iron uptake in Escherichia coli. J. Biochem. (Tokoy) 81, 749–756 (1977)

  16. Leive, L.: The barrier function of the gram-negative envelope. Ann. N.Y. Acad. Sci. 238, 109–129 (1974)

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

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

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

  20. Miller, J.H.: Experiments in molecular genetics. Cold Springer Harbor, New York: Cold Springer Harbor Laboratory 1972

  21. Nagel de Zwaig R., Zwaig, N., Istúriz, T., Sanchez, R.S.: Mutations affecting gluconate metabolism in Escherichia coli. J. Bact. 114, 463–468 (1973)

  22. Nakae, T.: Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. J. biol. Chem. 251, 2176–2178 (1976a)

  23. 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 (1976b)

  24. Nikaido, H.: Biosynthesis and assembly of lipopolysaccharide and the outer membrane layer of Gram-negative cell wall. In: Bacterial membranes and walls (Leive, L. ed.), pp. 131–208. New York: Marcel Dekker 1973

  25. Nikaido, H.: Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim. biophys. Acta (Amst.) 433, 118–132 (1976)

  26. Nikaido, H., Song, S.A., Shaltiel, L., Nurminen, M.: Outer membrane of Salmonella. XIV. Reduced transmembrane diffusion rates in porin-deficient mutants. Biochem. biophys. Res. Commun. 76, 324–330 (1977)

  27. Nurminen, M., Louatmaa, K., Sarvas, M., Mäkelä, P.H., Nakae, T.: Bacteriophage-resistant mutants of Salmonella typhimurium deficient in two major outer membrane proteins. J. Bact. 127, 941–955 (1976)

  28. O'Farrell, P.H.: High resolution two dimensional electrophoresis of proteins. J. biol. Chem. 250, 4007–4021 (1975)

  29. Pugsley, A.P., Reeves, P.: The role of colicin receptors in the uptake of ferrienterochelin by Escherichia coli K-12. Biochem. biophys. Res. Commun. 74, 903–911 (1977)

  30. Rosenbusch, J.P.: Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodesyl sulfate binding. J. biol. Chem. 249, 8019–8029 (1974)

  31. Sarma, V., Reeves, P.: Genetic locus (omB) affecting a major outer membrane protein in Escherichia coli K-12. J. Bact. 132, 23–27 (1977)

  32. Schleif, R.: Control of production of ribosomal protein. J. molec. Biol. 27, 41–55 (1967)

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

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

  35. Sekizawa, J., Inouye, S., Halegoua, S., Inouye, M.: Precursors of major outer membrane proteins of Escherichia coli. Biochem. biophys. Res. Commun. 77, 1126–1133 (1977)

  36. Smit, J., Kamio, Y., Nikaido, H.: Outer membrane of Salmonella typhimurium: Chemical analysis and freeze-fracture studies with lipopolysaccharide mutants. J. Bact. 124, 942–958 (1975)

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

  38. Wayne, R., Frick, K., Neilands, J.B.: Siderophore protection against colicins M, B, V, and Ia in Escherichia coli. J. Bact. 126, 7–12 (1977)

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

  40. Zimmermann, W., Rosselet, A.: The function of the outer membrane of Escherichia coli as a permeability barrier to β-lactam antibiotics. Antimicrob. Ag. Chemother. 12, 368–372 (1977)

Download references

Author information

Correspondence to Hiroshi Nikaido.

Additional information

This paper corresponds to paper XVI of the series dealing with the bacterial outer membrane from the laboratory of H.N. The preceding paper in the series is Nikaido, Bavoil, and Hirota, J. Bacteriol., in press

Communicated by W. Arber

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bavoil, P., Nikaido, H. & von Meyenburg, K. Pleiotropic transport mutants of Escherichia coli lack porin, a major outer membrane protein. Molec. Gen. Genet. 158, 23–33 (1977). https://doi.org/10.1007/BF00455116

Download citation

Keywords

  • Escherichia Coli
  • Permeability
  • Lactose
  • Outer Membrane
  • Wild Type Cell