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. 1978 Sep;135(3):1080–1090. doi: 10.1128/jb.135.3.1080-1090.1978

Porin activity in the osmotic shock fluid of Escherichia coli.

R Benz, B A Boehler-Kohler, R Dieterle, W Boos
PMCID: PMC222485  PMID: 357415

Abstract

Osmotic shock fluid of Escherichia coli exhibited pore-forming activity. This activity could be followed by an in vitro assay based on the conductivity increase for ions due to the presence of pores in black lipid membranes. The histogram (the distribution of conductivity increments in a single pore experiment) obtained with osmotic shock fluid from E. coli was identical to the histogram obtained by detergent-solubilized porin isolated from the outer membrane. The osmotic shock fluid from porin-negative mutants also exhibited pore activity, although the histogram and ion specificity were different from those of porin. Antibodies raised against detergent-solubilized porin were able to form precipitin lines by the Ouchterlony immunodiffusion technique when shock fluids, but not detergent-solubilized porin, were used. These antibodies prevented the formation of pores when shock fluids contained porin but not when shock fluids obtained from porin-negative mutants were used. Macroscopic membrane conductivity of shock fluids due to porin exhibited a concentration dependence, in contrast to detergent-solubilized porin. These results indicate that the hydrodynamic properties of periplasmic or "soluble" porin are different from those of the detergent-solubilized porin of the outer membrane. Periplasmic porin comprises about 0.7% of total protein in the osmotic shock fluid.

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Selected References

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  1. 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]
  2. Bassford P. J., Jr, Diedrich D. L., Schnaitman C. L., Reeves P. Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants. J Bacteriol. 1977 Aug;131(2):608–622. doi: 10.1128/jb.131.2.608-622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benz R., Stark G., Janko K., Läuger P. Valinomycin-mediated ion transport through neutral lipid membranes: influence of hydrocarbon chain length and temperature. J Membr Biol. 1973;14(4):339–364. doi: 10.1007/BF01868084. [DOI] [PubMed] [Google Scholar]
  4. Boos W. Bacterial transport. Annu Rev Biochem. 1974;43(0):123–146. doi: 10.1146/annurev.bi.43.070174.001011. [DOI] [PubMed] [Google Scholar]
  5. Boos W., Lengeler J., Hermann K. O., Unsöld H. J. The regulation of the beta-methylgalactoside transport system and of the galactose binding protein of Escherichia coli K12. Eur J Biochem. 1971 Apr 30;19(4):457–470. doi: 10.1111/j.1432-1033.1971.tb01336.x. [DOI] [PubMed] [Google Scholar]
  6. Bosch V., Braun V. Distribution of murein-lipoprotein between the cytoplasmic and outer membrane of Escherichia coli. FEBS Lett. 1973 Aug 15;34(2):307–310. doi: 10.1016/0014-5793(73)80818-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Braun V. Covalent lipoprotein from the outer membrane of Escherichia coli. Biochim Biophys Acta. 1975 Oct 31;415(3):335–377. doi: 10.1016/0304-4157(75)90013-1. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Chai T. J., Foulds J. Purification of protein A, an outer membrane component missing in Escherichia coli K-12 ompA mutants. Biochim Biophys Acta. 1977 Jul 22;493(1):210–215. doi: 10.1016/0005-2795(77)90274-4. [DOI] [PubMed] [Google Scholar]
  11. Costerton J. W., Ingram J. M., Cheng K. J. Structure and function of the cell envelope of gram-negative bacteria. Bacteriol Rev. 1974 Mar;38(1):87–110. doi: 10.1128/br.38.1.87-110.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Datta D. B., Arden B., Henning U. Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors. J Bacteriol. 1977 Sep;131(3):821–829. doi: 10.1128/jb.131.3.821-829.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dvorak H. F., Wetzel B. K., Heppel L. A. Biochemical and cytochemical evidence for the polar concentration of periplasmic enzymes in a "minicell" strain of Escherichia coli. J Bacteriol. 1970 Oct;104(1):543–548. doi: 10.1128/jb.104.1.543-548.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Endermann R., Krämer C., Henning U. Major outer membrane proteins of Escherichia coli K-12: evidence for protein II being a transmembrane protein. FEBS Lett. 1978 Feb 1;86(1):21–24. doi: 10.1016/0014-5793(78)80089-1. [DOI] [PubMed] [Google Scholar]
  15. Garrard W. T. Synthesis, assembly, and localization of periplasmic cytochrome c. J Biol Chem. 1972 Sep 25;247(18):5935–5943. [PubMed] [Google Scholar]
  16. Garten W., Henning U. Cell envelope and shape of Escherichia coli K12. Isolation and preliminary characterization of the major ghost-membrane proteins. Eur J Biochem. 1974 Sep 1;47(2):343–352. doi: 10.1111/j.1432-1033.1974.tb03699.x. [DOI] [PubMed] [Google Scholar]
  17. Garten W., Hindennach I., Henning U. The major proteins of the Escherichia coli outer cell envelope membrane. Characterization of proteins II* and III, comparison of all proteins. Eur J Biochem. 1975 Nov 1;59(1):215–221. doi: 10.1111/j.1432-1033.1975.tb02444.x. [DOI] [PubMed] [Google Scholar]
  18. Hasegawa Y., Yamada H., Mizushima S. Interactions of outer membrane proteins O-8 and O-9 with peptidoglycan sacculus of Escherichia coli K-12. J Biochem. 1976 Dec;80(6):1401–1409. doi: 10.1093/oxfordjournals.jbchem.a131413. [DOI] [PubMed] [Google Scholar]
  19. Henning U., Schmidmayr W., Hindennach I. Major proteins of the outer cell envelope membrane of Escherichia coli K-12: multiple species of protein I. Mol Gen Genet. 1977 Sep 9;154(3):293–298. doi: 10.1007/BF00571285. [DOI] [PubMed] [Google Scholar]
  20. Ichihara S., Mizushima S. Strain specificity of outer membrane proteins in Escherichia coli. J Biochem. 1977 May;81(5):1525–1530. [PubMed] [Google Scholar]
  21. Inouye M., Shaw J., Shen C. The assembly of a structural lipoprotein in the envelope of Escherichia coli. J Biol Chem. 1972 Dec 25;247(24):8154–8159. [PubMed] [Google Scholar]
  22. Jacobson G. R., Rosenbusch J. P. Abundance and membrane association of elongation factor Tu in E. coli. Nature. 1976 May 6;261(5555):23–26. doi: 10.1038/261023a0. [DOI] [PubMed] [Google Scholar]
  23. Johnson W. C., Silhavy T. J., Boos W. Two-dimensional polyacylamide gel electrophoresis of envelope proteins of Escherichia coli. Appl Microbiol. 1975 Mar;29(3):405–413. doi: 10.1128/am.29.3.405-413.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Kushnarev V. M., Smirnova T. A. Electron microscopy of alkaline phosphatase of Escherichia coli. Can J Microbiol. 1966 Aug;12(4):605–607. doi: 10.1139/m66-086. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Lee N., Scandella C., Inouye M. Spin labeling of a cysteine residue of the Escherichia coli outer membrane lipoprotein in its membrane environment. Proc Natl Acad Sci U S A. 1978 Jan;75(1):127–130. doi: 10.1073/pnas.75.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Livingston D. M. Immunoaffinity chromatography of proteins. Methods Enzymol. 1974;34:723–731. doi: 10.1016/s0076-6879(74)34094-3. [DOI] [PubMed] [Google Scholar]
  29. Lugtenberg B., Bronstein H., van Selm N., Peters R. Peptidoglycan-associated outer membrane proteins in gammegatine bacteria. Biochim Biophys Acta. 1977 Mar 17;465(3):571–578. doi: 10.1016/0005-2736(77)90274-7. [DOI] [PubMed] [Google Scholar]
  30. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  31. 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. 1976 Sep 23;147(3):251–262. doi: 10.1007/BF00582876. [DOI] [PubMed] [Google Scholar]
  32. MALAMY M., HORECKER B. L. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. doi: 10.1016/0006-291x(61)90020-1. [DOI] [PubMed] [Google Scholar]
  33. MacAlister T. J., Costerton J. W., Thompson L., Thompson J., Ingram J. M. Distribution of alkaline phosphatase within the periplasmic space of gram-negative bacteria. J Bacteriol. 1972 Sep;111(3):827–832. doi: 10.1128/jb.111.3.827-832.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. MacaAlister T. J., Irvin R. T., Costerton J. W. Cell surface-localized alkaline phosphatase of Escherichia coli as visualized by reaction product deposition and ferritin-labeled antibodies. J Bacteriol. 1977 Apr;130(1):318–328. doi: 10.1128/jb.130.1.318-328.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Nakae T. Outer membrane of Salmonella typhimurium: reconstitution of sucrose-permeable membrane vesicles. Biochem Biophys Res Commun. 1975 Jun 16;64(4):1224–1230. doi: 10.1016/0006-291x(75)90823-2. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Nakane P. K., Nichoalds G. E., Oxender D. L. Cellular localization of leucine-binding protein from Escherichia coli. Science. 1968 Jul 12;161(3837):182–183. doi: 10.1126/science.161.3837.182. [DOI] [PubMed] [Google Scholar]
  39. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  40. 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]
  41. Palva E. T., Randall L. L. Arrangement of protein I in Escherichia coli outer membrane: cross-linking study. J Bacteriol. 1978 Jan;133(1):279–286. doi: 10.1128/jb.133.1.279-286.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Palva E. T., Randall L. L. Nearest-neighbor analysis of Escherichia coli outer membrane proteins using cleavable cross-links. J Bacteriol. 1976 Sep;127(3):1558–1560. doi: 10.1128/jb.127.3.1558-1560.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pardee A. B., Watanabe K. Location of sulfate-binding protein in Salmonella typhimurium. J Bacteriol. 1968 Oct;96(4):1049–1054. doi: 10.1128/jb.96.4.1049-1054.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Randall-Hazelbauer L., Schwartz M. Isolation of the bacteriophage lambda receptor from Escherichia coli. J Bacteriol. 1973 Dec;116(3):1436–1446. doi: 10.1128/jb.116.3.1436-1446.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Randall L. L., Hardy S. J. Synthesis of exported proteins by membrane-bound polysomes from Escherichia coli. Eur J Biochem. 1977 May 2;75(1):43–53. doi: 10.1111/j.1432-1033.1977.tb11502.x. [DOI] [PubMed] [Google Scholar]
  46. Reithmeier R. A., Bragg P. D. Cross-linking of the proteins in the outer membrane of Escherichia coli. Biochim Biophys Acta. 1977 Apr 18;466(2):245–256. doi: 10.1016/0005-2736(77)90222-x. [DOI] [PubMed] [Google Scholar]
  47. 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]
  48. Salton M. R., Owen P. Bacterial membrane structure. Annu Rev Microbiol. 1976;30:451–482. doi: 10.1146/annurev.mi.30.100176.002315. [DOI] [PubMed] [Google Scholar]
  49. Schmitges C. J., Henning U. The major proteins of the Escherichia coli outer cell-envelope membrane. Heterogeneity of protein I. Eur J Biochem. 1976 Mar 16;63(1):47–52. doi: 10.1111/j.1432-1033.1976.tb10205.x. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Schumacher G., Bussmann K. Cell-free synthesis of proteins related to sn-glycerol-3-phosphate transport in Escherichia coli. J Bacteriol. 1978 Jul;135(1):239–250. doi: 10.1128/jb.135.1.239-250.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Silhavy T. J., Casadaban M. J., Shuman H. A., Beckwith J. R. Conversion of beta-galactosidase to a membrane-bound state by gene fusion. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3423–3427. doi: 10.1073/pnas.73.10.3423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Smith W. P., Tai P. C., Thompson R. C., Davis B. D. Extracellular labeling of nascent polypeptides traversing the membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2830–2834. doi: 10.1073/pnas.74.7.2830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Steven A. C., Heggeler B., Müller R., Kistler J., Rosenbusch J. P. Ultrastructure of a periodic protein layer in the outer membrane of Escherichia coli. J Cell Biol. 1977 Feb;72(2):292–301. doi: 10.1083/jcb.72.2.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Szmelcman S., Hofnung M. Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor. J Bacteriol. 1975 Oct;124(1):112–118. doi: 10.1128/jb.124.1.112-118.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Szmelcman S., Schwartz M., Silhavy T. J., Boos W. Maltose transport in Escherichia coli K12. A comparison of transport kinetics in wild-type and lambda-resistant mutants as measured by fluorescence quenching. Eur J Biochem. 1976 May 17;65(1):13–19. doi: 10.1111/j.1432-1033.1976.tb10383.x. [DOI] [PubMed] [Google Scholar]
  58. Uemura J., Mizushima S. Isolation of outer membrane proteins of Escherchia coli and their characterization on polyacrylamide gel. Biochim Biophys Acta. 1975 Dec 1;413(2):163–176. doi: 10.1016/0005-2736(75)90101-7. [DOI] [PubMed] [Google Scholar]
  59. Yu F., Mizushima S. Stimulation by lipopolysaccharide of the binding of outer membrane proteins O-8 and O-9 to the peptidoglycan layer of Escherichia coli K--12. Biochem Biophys Res Commun. 1977 Feb 21;74(4):1397–1402. doi: 10.1016/0006-291x(77)90597-6. [DOI] [PubMed] [Google Scholar]
  60. von Meyenburg K., Nikaido H. Outer membrane of gram-negative bacteria. XVII. Secificity of transport process catalyzed by the lambda-receptor protein in Escherichia coli. Biochem Biophys Res Commun. 1977 Oct 10;78(3):1100–1107. doi: 10.1016/0006-291x(77)90534-4. [DOI] [PubMed] [Google Scholar]

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