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Volume 142, Issue 8

Protein C (OprC) of the outer membrane of is a copper-regulated channel protein Free

Abstract

Protein C (OprC) of the outer membrane of forms small channels, as assayed by the liposome swelling method. We report here that OprC functions as a channel-forming and copper-binding protein. OprC purified to homogeneity formed a channel in planar lipid bilayers with an ion conductance of about 200 pS in 1 M NaCl. Cloning and sequencing of the gene encoding OprC revealed that it specified a polypeptide comprising 723 and 668 amino acid residues for the precursor and mature polypeptides ( 73372), respectively. The amino acid sequence of OprC showed the highest degree of similarity with that of NosA of (65% sequence identity) which conveys Cu to intracellular acceptor(s). OprC showed high copper-binding activity ( = 2·6 μM) in aqueous solution containing surfactant. The expression of OprC appeared to be repressed by exogenous Cu and derepressed by anaerobiosis in the presence of nitrate. These results suggest that OprC might be involved in copper utilization.

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Keyword(s): copper-binding protein , outer membrane , porin , protein C (OprC) and Pseudomonas aeruginosa
© Society for General Microbiology 1996
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1996-08-01
2025-11-10

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References

  1. Adler J., Hazelbauer G.L., Dahl M.M. 1973; Chemotaxis toward sugars in Escherichia coli . J Bacterial 115:824–847
     [Google Scholar]
  2. Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410
     [Google Scholar]
  3. Benz R., Janko K., Boos W., Läuger P. 1978; Formation of large, ion-permeable membrane channels by the matrix protein (porin) of Escherichia coli . Biochim Biophys Acta 511:305–319
     [Google Scholar]
  4. Benz R., Schmid A., Nakae T., Vos-Scheperkeuter G.H. 1986; Pore formation by LamB of Escherichia coli in lipid bilayer membranes. J Bacteriol 165:978–986
     [Google Scholar]
  5. Bitter W., Marugg J.D., de Weger L.A., Tommassen J., Weisbeek P.J. 1991; The ferric-pseudobactin receptor PupA of Pseudomonas putida WCS358: homology to TonB-dependent Escherichia coli receptors and specificity of the protein. Mol Microbiol 5:647–655
     [Google Scholar]
  6. Bischer K.H., Cullmann W., Dick W., Wendt S., Opferkuch W. 1987; Imipenem resistance in Pseudomonas aeruginosa is due to diminished expression of outer membrane proteins. J Infect Dis 156:681–684
     [Google Scholar]
  7. Cha J., Cooksey D.A. 1991; Copper resistance in Pseudomonas syringae mediated by periplasmic and outer membrane proteins. Proc Natl Acad Sci USA 888915–8919
     [Google Scholar]
  8. Dean C.R., Poole K. 1993; Cloning and characterization of the ferric enterobactin receptor gene (PfeA) of Pseudomonas aeruginosa . J Bacteriol 175:317–324
     [Google Scholar]
  9. Deretic V., Konyecsni W.M., Mohr C.D., Martin D.W., Hibler N.S. 1989; Common denominators of promoter control in Pseudomonas and other bacteria. Bio/Technology 7:1249–1254
     [Google Scholar]
  10. Doige C., Ames G.F.L. 1993; ATP-dependent transport systems in bacteria and humans: relevance to cystic fibrosis and multidrug resistance. Annu Ren Microbiol 47:291–319
     [Google Scholar]
  11. Galimand M., Gamper M., Zimmermann A., Haas D. 1991; Positive FNR-like control of anaerobic arginine degradation and nitrate respiration in Pseudomonas aeruginosa . J Bacteriol 173:1598–1606
     [Google Scholar]
  12. Heller K., Kadner R.J. 1985; Nucleotide sequence of the gene for the vitamin B12 receptor protein in the outer membrane of Escherichia coli . J Bacteriol 161:904–908
     [Google Scholar]
  13. Hirano H., Watanabe T. 1990; Microsequencing of proteins electrotransferred onto immobilizing matrices from polyacrylamide gel electrophoresis: application to an insoluble protein. Electrophoresis 11:573–580
     [Google Scholar]
  14. Hirayama H. 1976; Spectrophotometric determination of trace amounts of copper based on its catalytic oxidation of o-amino-phenol. J Chem Soc Jpn 5:768–772
     [Google Scholar]
  15. Ishii J., Nakae T. 1993; Lipopolysaccharide promoted opening of the porin channel. FEBS Lett 320:251–255
     [Google Scholar]
  16. Küpper H., Sekiya T., Rosenberg M., Egan J., Landy A. 1978; A rho-dependent termination site in the gene coding for tyrosine tRNA su3 of Escherichia coli . Nature 272:423–428
     [Google Scholar]
  17. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
     [Google Scholar]
  18. Lee H.S., Hancock R.E.W., Ingraham J.L. 1989; Properties of a Pseudomonas stutszeri outer membrane channel-forming protein (NosA) required for production of copper-containing N2O reductase. J Bacteriol 171:2096–2100
     [Google Scholar]
  19. Lee H.S., Abdelal A.H.T., Clark M.A., Ingraham J.L. 1991; Molecular characterization of nos A, a Pseudomonas stutizeri gene encoding an outer membrane protein required to make copper- containing NaO reductase. J Bacteriol 173:5406–5413
     [Google Scholar]
  20. Lipman D.J., Pearson W.R. 1985; Rapid and sensitive protein similarity searches. Science 227:1435–1441
     [Google Scholar]
  21. Lundrlgan M.D., Kadner R.J. 1986; Nucleotide sequence of the gene for the ferrienterochelin receptor FepA in Escherichia coli : homology among outer membrane receptors that interact with TonB. J Biol Chem 261:10797–10801
     [Google Scholar]
  22. Mokhele K., Tang Y.J., Clark M.A., Ingraham J.L. 1987; A Pseudomonas stutgeri outer membrane protein inserts copper into N2O reductase. J Bacteriol 169:5721–5726
     [Google Scholar]
  23. Nau C.D., Konisky J. 1989; Evolutionary relationship between the TonB-dependent outer membrane transport proteins: nucleotide and amino acid sequences of the Escherichia coli Colicin I receptor gene. J Bacteriol 171:1041–1047
     [Google Scholar]
  24. Obara M., Nakae T. 1992; Porin of Pseudomonas aeruginosa forms low conductance ion channel in planar lipid bilayers. Biochem Biophys Rfj Commun 186:645–651
     [Google Scholar]
  25. Pearson W.R., Lipman D.J. 1988; Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 852444–2448
     [Google Scholar]
  26. Quinn J.P., Dudek E.J., DiVincenzo C.A., Lucks D.A., Lerner S.A. 1986; Emergence of resistance to imipenem during therapy for Pseudomonas aeruginosa infections. J Infect Dis 154:289–294
     [Google Scholar]
  27. Rosenberg M., Court D. 1979; Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Ren Genet 13:319–335
     [Google Scholar]
  28. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 745463–5467
     [Google Scholar]
  30. Satake S., Yoshihara E., Nakae T. 1990; Diffusion of β-lactam antibiotics through liposome membrane reconstituted from purified porins of the outer membrane of Pseudomonas aeruginosa . Antimicrob Agents Chemother 34:685–690
     [Google Scholar]
  31. Sauer M., Hantke K., Braun V. 1990; Sequence of the fhuE outer-membrane receptor gene of Escherichia coli K12 and properties of mutants. Mol Microbiol 4:427–437
     [Google Scholar]
  32. Skare J.T., Ahmer B.M.M., Seachord C.L., Darveau R.P., Postle K. 1993; Energy transduction between membranes. TonB, a cytoplasmic membrane protein, can be chemically cross-linked in vivo to the outer membrane receptor FepA. J Biol Chem 268:16302–16308
     [Google Scholar]
  33. Solioz M., Odermatt A. 1995; Copper and silver transport by CopB-ATPase in membrane vesicles of Enterococcus hirae . J Biol Chem 270:9217–9221
     [Google Scholar]
  34. Toschka H.Y., Höpfl P., Ludwig W., Schleifer K.H., Ulbrich N., Erdmann V.A. 1988; Complete nucleotide sequence of a 16S ribosomal RNA gene from Pseudomonas aeruginosa . Nucleic Acids Res 16:2348
     [Google Scholar]
  35. Trias J., Nikaido H. 1990; Outer membrane protein D2 catalyzes facilitated diffusion of carbapenems and penems through the outer membrane of Pseudomonas aeruginosa . Antimicrob Agents Chemother 34:52–57
     [Google Scholar]
  36. Yamano Y., Nishikawa T., Komatsu Y. 1993; Cloning and nucleotide sequence of anaerobically induced porin protein El (OprE) of Pseudomonas aeruginosa PAOl. Mol Microbiol 8:993–1004
     [Google Scholar]
  37. Yoneyama H., Nakae T. 1986; A small diffusion pore in the outer membrane of Pseudomonas aeruginosa . Eur J Biochem 157:33–38
     [Google Scholar]
  38. Yoneyama H., Nakae T. 1991; Cloning of the protein D2 gene of Pseudomonas aeruginosa and its functional expression in the imipenem-resistant host. FEBS Lett 283:177–179
     [Google Scholar]
  39. Yoneyama H., Akatsuka A., Nakae T. 1986; The outer membrane of Pseudomonas aeruginosa is a barrier against the penetration of disaccharides. Biochem Biophys Res Commun 134:106–112
     [Google Scholar]
  40. Yoneyama H., Yoshihara E., Nakae T. 1992; Nucleotide sequence of the protein D2 gene of Pseudomonas aeruginosa . Antimicrob Agents Chemother 36:1791–1793
     [Google Scholar]
  41. Yoshihara E., Nakae T. 1989; Identification of porins in the outer membrane of Pseudomonas aeruginosa that form small diffusion pores. J Biol Chem 264:6297–6301
     [Google Scholar]
  42. Yoshihara E., Nakae T. 1992; Separation of gate- and channelforming domains in the pore-forming protein of the outer membrane of Pseudomonas aeruginosa . FEBS Lett 306:5–8
     [Google Scholar]
  43. Yoshihara E., Yoneyama H., Nakae T. 1991; In vitro assembly of the functional porin trimer from dissociated monomers in Pseudomonas aeruginosa . J Biol Chem 266:952–957
     [Google Scholar]
  44. Zannoni D. 1989; The respiratory chains of pathogenic pseudomonads. Biochim Biophys Acta 975:299–316
     [Google Scholar]
/content/journal/micro/10.1099/13500872-142-8-2137
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Protein C (OprC) of the outer membrane of Pseudomonas aeruginosa is a copper-regulated channel protein
Microbiology 142, 2137 (1996); https://doi.org/10.1099/13500872-142-8-2137
/content/journal/micro/10.1099/13500872-142-8-2137
/content/journal/micro/10.1099/13500872-142-8-2137
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