1887

Microbiology Society logo

Volume 140, Issue 12

(formerly ) porin is an oligomer composed of two component proteins Free

Abstract

The 81 kDa protein (designated OpcPO) which forms a diffusion pore in the outer membrane of (formerly ) has a unique characteristic in that when the purified protein is heated it yields a major 36 kDa protein (designated OpcP1) and a minor 27 kDa protein (designated OpcP2). Moreover, incubation of OpcPO in citrate buffer at pH 3.0 produced an unusual dissociation into 72 kDa and 27 kDa proteins. For the characterization of OpcPO and its derivatives, OpcP1 and OpcP2 from purified OpcPO were isolated by preparative SDS-PAGE. Reconstitution of OpcPO using purified preparations of OpcP1 and OpcP2 indicated that these derivatives were not proteolytic fragments of OpcPO. Moreover, immunoblot assays with murine polyclonal antisera specific for OpcP1 and OpcP2 yielded the following results: (i) OpcP1 and OpcP2 are immunologically distinguishable proteins; (ii) the unusual dissociation of OpcPO in citrate buffer at pH 3.0 resulted in the release of OpcP2 from OpcPO, and the resulting 72 kDa protein was probably an oligomer of OpcP1; (iii) purified OpcP1 itself produced two additional 53 kDa and 72 kDa proteins spontaneously following elution from the bottom of the SDS-PAGE gel. From these findings, it was concluded that OpcPO is formed by the non-covalent association of OpcP2 with an oligomer of OpcP1 that has the ability to self-assemble.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Burkholderia (Pseudomonas) cepacia , outer membrane protein , porins and self assembly
© Society for General Microbiology, 1994
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-12-3285
1994-12-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/12/mic-140-12-3285.html?itemId=/content/journal/micro/10.1099/13500872-140-12-3285&mimeType=html&fmt=ahah

References

  1. Anwar H., Brown M.R.W., Cozens R.M., Lambert P.A. Isolation and characterization of the outer and cytoplasmic membranes of Pseudomonas cepacia. J Gen Microbiol 1983; 129:499–507
     [Google Scholar]
  2. Aronoff A.C. Outer membrane permeability in Pseudomonas cepacia: diminished porin content in a z-lactam-resistant mutant and in resistant cystic fibrosis isolates. Antimicrob Agents Chemother 1988; 32:1636–1639
     [Google Scholar]
  3. Aronoff S.C., Stern R.C. Serum IgG antibody to outer membrane antigens of Pseudomonas cepacia and Pseudomonas aeruginosa in cystic fibrosis. J Infect Dis 1988; 157:934–940
     [Google Scholar]
  4. Benz R. Structure and function of porins from Gramnegative bacteria. Annu Rep Microbiol 1988; 42:359–393
     [Google Scholar]
  5. Burns J.L., Clark D.K. Salicylate-inducible antibiotic resistance in Pseudomonas cepacia associated with absence of a pore-forming outer membrane protein. Antimicrob Agents Chemother 1992; 36:2280–2285
     [Google Scholar]
  6. Ederer G.M., Matsen J.M. Colonization and infection with Pseudomonas cepacia. J Infect Dis 1972; 125:613–618
     [Google Scholar]
  7. Fass R.J., Barnishan J. In vitro susceptibilities of nonfermentative Gram-negative bacilli other than Pseudomonas aeruginosa to 32 antimicrobial agents. Rev Infect Dis 1980; 2:841–853
     [Google Scholar]
  8. Gilligan P.H. Microbiology of airway disease in patients with cystic fibrosis. Clin Microbiol Rev 1991; 4:35–51
     [Google Scholar]
  9. Hancock R.E.W. Role of porins in outer membrane permeability. J Bacterial 1987; 169:929–933
     [Google Scholar]
  10. Hancock R.E.W., Carey A.M. Outer membrane of Pseudomonas aeruginosa: heat-and 2-mercaptoethanol-modifiable proteins. J Bacteriol 1979; 140:902–910
     [Google Scholar]
  11. Harlow E., Lane D. Antibodies: A Laboratory Manual 1988 Cold Spring Harbor, New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  12. Kameyama K., Nakae T., Takagi T. Estimation of molecular weight of membrane proteins in the presence of SDS by low-angle laser light scattering combined with high-performance porous silica gel chromatography confirmation of the trimer structure of porin of the Escherichia coli outer membrane. Biochim Biophys Acta 1982; 706:19–26
     [Google Scholar]
  13. Kropp H., Gerckeus L., Sundelof J.G., Kahan F.M. Antibacterial activity of imipenem: the first thienamycin antibiotic. Rev Infect Dis 1985; 7:S389–410
     [Google Scholar]
  14. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
     [Google Scholar]
  15. Maezawa S., Hayashi Y., Nakae T., Ishii J., Kameyama K., Takagi T. Determination of molecular weight of membrane proteins by the use of low-angle laser light scattering combined with high-performance gel chromatography in the presence of a non-ionic surfactant. Biochim Biophys Acta 1983; 747:291–297
     [Google Scholar]
  16. Nakae T. Outer membrane of Salmonella typhimurium: Reconstitution of sucrose-permeable membrane vesicles. Biochem Biophys Res Commun 1975; 64:1224–1230
     [Google Scholar]
  17. Nakae T. Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. J Biol Chem 1976; 251:2176–2178
     [Google Scholar]
  18. Nakae T. Outer-membrane permeability of bacteria. Crit Rep Microbiol 1986; 13:1–62
     [Google Scholar]
  19. Nakae T., Ishii J., Tokunaga M. Subunit structure of functional porin oligomers that form permeability channels in the outer membrane of Escherichia coli. J Biol Chem 1979; 254:1457–1461
     [Google Scholar]
  20. Nikaido H. Outer membrane barrier as a mechanism of antimicrobial resistance. Antimicrob Agents Chemother 1989; 33:1831–1836
     [Google Scholar]
  21. Nikaido H., Red J. Biogenesis of prokaryotic pores. Experientia 1990; 46:174–180
     [Google Scholar]
  22. Nikaido H., Rosenberg E.Y. Porin channels in Escherichia coli: Studies with liposomes reconstituted from purified proteins. J Bacteriol 1983; 153:241–252
     [Google Scholar]
  23. Nikaido H., Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rep 1985; 49:1–32
     [Google Scholar]
  24. Palva E.T., Randall L.L. Arrangement of protein I in Escherichia coli outer membrane: Cross-linking study. T Bacteriol 1978; 133:279–286
     [Google Scholar]
  25. Palva E.T. Arrangement of protein II, a phage-directed major outer membrane protein in Escherichia coli. FEMS Microbiol Lett 1979; 5:73–76
     [Google Scholar]
  26. Parr T.R., Moore R.A., Moore L.V., Hancock R.E.W. Role of porins in intrinsic antibiotic resistance of Pseudomonas cepacia. Antimicrob Agents Chemother 1987; 31:121–123
     [Google Scholar]
  27. Philips I., Eykyn S. Pseudomonas cepacia (multivorans) septicemia in an intensive care unit. Lancet 1971; 1:375–377
     [Google Scholar]
  28. Rosenbusch J.P. Structure and functional properties of porin channels in E. coli outer membranes. Experientia 1990; 46:167–173
     [Google Scholar]
  29. Smith D.L., Gumery L.B., Smith E.G., Stableforth D.E., Kaufmann M.E., Pitt T.L. Epidemie of Pseudomonas cepacia in an adult cystic fibrosis unit: evidence of person-to-person transmission. J Clin Microbiol 1993; 31:3017–3022
     [Google Scholar]
  30. Steven A.C., Ten Heggler B., Muller R., Kistler J., Rosenbusch J.P. Ultrastructure of a periodic protein layer in the outer membrane of Escherichia coli. J Cell Biol 1977; 72:292–301
     [Google Scholar]
  31. Tokunaga M., Tokunaga H., Okajima Y., Nakae T. Characterization of porin from the outer membrane of Salmonella typhimurium 2. Physical properties of the functional oligomeric aggregates. Eur J Biochem 1979; 95:441–448
     [Google Scholar]
  32. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sei USA 1979; 76:4350–4354
     [Google Scholar]
  33. Yoshimura F., Nikaido H. The porin channels of Escherichia coli K-12. Antimicrob Agents Chemother 1985; 27:84–92
     [Google Scholar]
  34. Yu J., Ichihara S., Mizushima S. A major outer membrane protein (0-8) of Escherichia coli K-12 exists as a trimer in sodium dodecylsulfate solution. FEBS Lett 1979; 100:71–74
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-12-3285
Loading
Burkholderia(formerly Pseudomonas) cepaciaporin is an oligomer composed of two component proteins
Microbiology 140, 3285 (1994); https://doi.org/10.1099/13500872-140-12-3285
/content/journal/micro/10.1099/13500872-140-12-3285
/content/journal/micro/10.1099/13500872-140-12-3285
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error