1887

Abstract

N-terminal sequence analysis of peptides generated by proteolytic treatment of the OE 28.3 major outer-membrane protein OprF, embedded in outer membranes or present in whole cells, indicated a surface-exposed location for the proline-rich region of the protein. This region is absent from the and OprFs. Evidence was obtained for the presence of additional exposed but less accessible regions in the carboxy half of OprF. Four OprF-specific monoclonal antibodies were all directed to the C-terminal part of the protein but did not recognize a surface-exposed epitope as shown by flow cytometry. Our data support the model previously proposed for OprF in which the entire protein is embedded in the outer membrane, unlike the topology proposed for the major outer-membrane protein from , OmpA, whose carboxy half resides in the periplasmic space. For six other strains producing OprF proteins with different isoelectric points, the primary structure was determined by sequence analysis of the PCR-amplified genes. The proline-rich domain represented the most conserved region of the different OprFs. Based on the sequence of its gene, it was shown that the mushroom pathogen is quite closely related to . Comparative sequence analysis further showed that the carboxy half of OprF contains a sequence motif that is well conserved in the enterobacterial OmpA proteins but is also present in a number of other outer-membrane proteins, including peptidoglycan-associated lipoproteins.

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1994-06-01
2021-05-16
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References

  1. Arnheim N., Erlich H. 1992; Polymerase chain reaction strategy. Annu Rev Biochem 61:131–156
    [Google Scholar]
  2. Bellido F., Martin N. L., Siehnel R. J., Hancock R. E. W. 1992; Reevaluation, using intact cells, of the exclusion limit and role of porin Opr F in Pseudomonas aeruginosa outer membrane permeability. J Bacteriol 174:5196–5203
    [Google Scholar]
  3. Butt N. J., Virji M., Vayreda F., Lambden P. R., Heckels I. E. 1990; Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by tvpe-specific and cross-reacting monoclonal antibodies. J Gen Microbiol 136:2165–2172
    [Google Scholar]
  4. Bystrom A. S., Hjalmarsson K. J., Wikstrom P. M., Bjork G. R. 1983; The nucleotide sequence of an Escherichia coli operon containing genes for the tRNA (nAG) methyltransferase, the ribosomal proteins SI6 and LI 9 and a 21-K polypeptide. EMBO J 2:899–905
    [Google Scholar]
  5. Charbit A., Ronco J., Michel V., Werts C., Hofnung M. 1991; Permissive sites and topology of an outer membrane protein with a reporter epitope. J Bacteriol 173:262–275
    [Google Scholar]
  6. Chen R., Henning U. 1987; Nucleotide sequence of the gene for the peptidoglycan-associated lipoprotein of Escherichia coli K12. Eur J Biochem 163:73–77
    [Google Scholar]
  7. Dean D., Schachter J., Dawson C. R., Stephens R. S. 1992; Comparison of the major outer membrane protein variant sequence regions of B/Ba isolates: a molecular epidemiologic approach to Chlamydia trachomatis infections. J Infect Dis 166:383–392
    [Google Scholar]
  8. Deich R. A., Metcalf B. J., Finn C. W., Farley J. E., Green B. A. 1988; Cloning of genes encoding a 15000-dalton peptidoglycan- associated outer membrane lipoprotein and an antigenically related 15000-dalton protein from Haemophilus influenzae. J Bacteriol 170:489–498
    [Google Scholar]
  9. De Mot R., Vanderleyden J. 1989; Application of two-dimensional protein analysis for strain fingerprinting and mutant analysis of Azpspirillum species. Can J Microbiol 35:960–967
    [Google Scholar]
  10. De Mot R., Vanderleyden J. 1991; Purification of a root- adhesive outer membrane protein of root-colonizing Pseudomonas fluorescens. FEMS Microbiol Eett 81:323–328
    [Google Scholar]
  11. De Mot R., Proost P., Van Damme J., Vanderleyden J. 1992; Homology of the root adhesin of Pseudomonas fluorescens OE 28.3 with porin F of P. aeruginosa and P. syringae. Mol and Gen Genet 231:489–493
    [Google Scholar]
  12. De Mot R., Laeremans T., Schoofs G., Vanderleyden J. 1993; Characterization of the rec A gene from Pseudomonas fluorescens OE 28.3 and construction of a rec A mutant. J Gen Microbiol 139:49–57
    [Google Scholar]
  13. Duchene M., Schweizer A., Lottspeich F., Krauss G., Marget M., Vogel K., von Specht B.-U., Domdey H. 1988; Sequence and transcriptional start site of the Pseudomonas aeruginosa outer membrane porin protein F gene. J Bacteriol 170:155–162
    [Google Scholar]
  14. Engleberg N. C., Howe D. C., Rogers J. E., Arroyo J, Eisenstein B. I. 1991; Characterization of a Eegionella pneumophilia gene encoding a lipoprotein antigen. Mol Microbiol 5:2021–2029
    [Google Scholar]
  15. Finnen R. L., Martin N. L., Siehnel R. J., Woodruff W. A., Rosok M., Hancock R. E. W. 1992; Analysis of the Pseudomonas aeruginosa major outer membrane protein Opr F by use of truncated Opr F derivatives and monoclonal antibodies. J Bacteriol 174:4977–4985
    [Google Scholar]
  16. Gentry-Weeks C.R., Hultsch A.-L., Kelly S. M., Keith J. M., Curtiss R. 1992; Cloning and sequencing of a gene encoding a 21-kilodalton outer membrane protein from Bordetella avium and expression of the gene in Salmonella typhimurium. J Bacteriol 174:7729–7742
    [Google Scholar]
  17. Georgiou G., Poetschke H. L., Stathopoulos C., Francisco J. A. 1993; Practical applications of engineering Gram-negative bacterial surfaces. Trends Biotechnol 11:6–10
    [Google Scholar]
  18. Goor M., Vantomme R., Swings J., Gillis M., Kersters K., De Ley J. 1986; Phenotypic and genotypic diversity of Pseudomonas tolaasii and white line reacting organisms isolated from cultivated mushrooms. J Gen Microbiol 132:2249–2264
    [Google Scholar]
  19. Gotoh N., Wakebe H., Nishino T. 1989a; Ultrastructural aspects of fragility of Pseudomonas aeruginosa outer membrane devoid of protein F. FEMS Microbiol Eett 59:51–54
    [Google Scholar]
  20. Gotoh N., Wakebe H., Yoshihara E., Nakae T., Nishino T. 1989b; Role of protein F in maintaining structural integrity of the Pseudomonas aeruginosa outer membrane. J Bacteriol 171:983–990
    [Google Scholar]
  21. Gotschlich E. C., Seiff M., Blake M. S. 1987; The DNA sequence of the structural gene of gonococcal protein III and the flanking region containing a repetitive sequence. Homology of protein III with enterobacterial Omp A proteins. J Exp Med 165:471–482
    [Google Scholar]
  22. Harlowe E., Lane D. 1988 Antibodies. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. Hayes L. J., Pickett M. A., Conlan J. W., Ferris S., Everson J. S., Ward M. E., Clarke I. N. 1990; The major outer-membrane proteins of Chlamydia trachomatis serovars A and B: intra-serovar amino acid changes do not alter specificities of serovar- and C subspecies-reactive antibody-binding domains. J Gen Microbiol 136:1559–1566
    [Google Scholar]
  24. Higgins D.G., Sharp P. M. 1988; Clustal: a package for performing multiple sequence alignment on a microcomputer. Gene 73:237–244
    [Google Scholar]
  25. Hughes E. E., Gilleland L. B., Gilleland H. E. 1992; Synthetic peptides representing epitopes of outer membrane protein F of Pseudomonas aeruginosa that elicit antibodies reactive with whole cells of heterologous immunotype strains of P. aeruginosa. Infect Immun 60:3497–3503
    [Google Scholar]
  26. Kaltenboeck B., Kousoulas K. G., Storz J. 1993; Structures and allelic diversity and relationships among the major outer membrane protein (omp A) genes of the four chlamydial species. J Bacteriol 175:487–502
    [Google Scholar]
  27. Klose M., MacIntyre S., Schwarz H., Henning U. 1988a; The influence of amino substitutions within the mature part of an Escherichia coli outer membrane protein (Omp A) on assembly of the polypeptide into its membrane. J Biol Chem 263:13297–13302
    [Google Scholar]
  28. Klose M., Schwarz H., MacIntyre S., Freudl R., Eschbach M.-L., Henning U. 1988b; Internal deletions in the gene for an Escherichia coli outer membrane protein define an area possibly important for recognition of the outer membrane by this polypeptide. J Biol Chem 263:13291–13296
    [Google Scholar]
  29. Klugman K. P., Gotschlich E. C., Blake M. S. 1989; Sequence of the structural gene (rmp M) for the class 4 outer membrane protein of Neisseria meningitidis, homology of the protein to gonococcal protein III and Escherichia coli Omp A, and construction of meningococcal strains that lack class 4 protein. Infect Immun 57:2066–2071
    [Google Scholar]
  30. Lampe M. F., Suchland R. J., Stamm W. E. 1993; Nucleotide sequence of the variable domains within the major outer membrane protein gene from serovariants of Chlamydia trachomatis. Infect Immun 61:213–219
    [Google Scholar]
  31. Lawrence J. G., Ochman H., Hartl D. L. 1991; Molecular and evolutionary relationships among enteric bacteria. J Gen Microbiol 137:1911–1921
    [Google Scholar]
  32. Little M., Fuchs P., Breitling F., Diibel S. 1993; Bacterial surface presentation of proteins and peptides: an alternative to phage technology. Trends Biotechnol 11:3–5
    [Google Scholar]
  33. Lugtenberg B.J.J., de Weger L.A., Bennett J. W. 1991; Microbial stimulation of plant growth and protection from disease. Curr Opin Biotechnol 2:457–464
    [Google Scholar]
  34. Maiden M.C.J., Suker J., McKenna A.J., Bygraves J. A., Feavers I. M. 1991; Comparison of the class 1 outer membrane proteins of eight serological reference strains of Neisseria meningitidis. Mol Microbiol 5:727–736
    [Google Scholar]
  35. Matsudaira P.T. 1989 A Practical Guide to Protein and Peptide Purification for Microsequencing. San Diego: Academic Press;
    [Google Scholar]
  36. Mazzola M., Cook R. J., Tomashow L. S., Weller D. M., Pierson L. S. 1992; Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent Pseudomonads in soil habitats. Appl Environ Microbiol 58:2616–2624
    [Google Scholar]
  37. Morona R., Klose M., Henning U. 1984; Escherichia coli K-12 outer membrane protein (Omp A) as a bacteriophage receptor: analysis of mutant genes expressing altered proteins. J Bacteriol 159:570–578
    [Google Scholar]
  38. Munson R. S., Grass S., West R. 1993; Molecular cloning and sequence of the gene for outer membrane protein P5 of Haemophilus influenzae. Infect Immun 61:4017–4020
    [Google Scholar]
  39. Murphy T. F., Kirkham C., Lesse A. J. 1993; The major heat- modifiable outer membrane protein CD is highly conserved among strains of Branhamella catarrhalis. Mol Microbiol 10:87–97
    [Google Scholar]
  40. Mutharia L.M., Hancock R. E. W. 1985; Characterization of two surface-localized antigenic sites on porin protein F of Pseudomonas aeruginosa. Can J Microbiol 31:381–386
    [Google Scholar]
  41. Nelson M. B., Apicella M. A., Murphy T. F., Vankeulen H., Spotila L. D., Rekosch D. 1988; Cloning and sequencing of Haemophilus influenzae outer membrane protein P6. Infect Immun 56:128–134
    [Google Scholar]
  42. Nikaido H., Nikaido K., Harayama S. 1991; Identification and characterization of porins in Pseudomonas aeruginosa. J Biol Chem 266:770–779
    [Google Scholar]
  43. O'Sullivan D.J., O'Gara F. 1992; Traits of fluorescent Pseudomonas spp. involved in suppression of plant root pathogens. Microbiol Rev 50:662–676
    [Google Scholar]
  44. Rainey P. B., Brodey C. L., Johnstone K. 1992; Biology of Pseudomonas tolaasii, cause of brown blotch disease of the cultivated mushroom. Adv Plant Pathol 8:95–117
    [Google Scholar]
  45. Srikumar R., Chin A. C., Vachon V., Richardson C. D., Ratdiffe M. J. H., Saarinen L., Kayhty H., Makela P. H., Coulton J. W. 1992; Monoclonal antibodies specific to porin of Haemophilus influenzae type b: localization of their cognate epitopes and tests of their biological activities. Mol Microbiol 6:665–676
    [Google Scholar]
  46. Stead D.E. 1992; Grouping of plant-pathogenic and some other Pseudomonas spp. by using cellular fatty acid profiles. Int J Syst Bacteriol 42:281–295
    [Google Scholar]
  47. Sugawara E., Nikaido H. 1992; Pore-forming activity of Omp A protein of Escherichia coli. J Biol Chem 267:2507–2511
    [Google Scholar]
  48. Surrey T., Jahnig F. 1992; Refolding and oriented insertion of a membrane protein into a lipid bilayer. Proc Natl Acad Sci USA 89:7457–7461
    [Google Scholar]
  49. Szwajcer-Dey E, Rasmussen J., Meldal M., Breddam K. 1992; Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp. J Bacteriol 174:2454–2459
    [Google Scholar]
  50. Ullstrom C. A., Siehnel R., Woodruff W., Steinbach S., Hancock R. E. W. 1991; Conservation of the gene for outer membrane protein Opr F in the family Pseudomonadaceae-. sequence of the Pseudomonas syringae opr F gene. J Bacteriol 173:768–775
    [Google Scholar]
  51. Vlassak K., Van Holm L., Duchateau L., Vanderleyden J., De Mot R. 1992; Isolation and characterization of fluorescent Pseudomonas associated with the roots of rice and banana grown in Sri Lanka. Plant Soil 145:51–63
    [Google Scholar]
  52. Vogel H., Jahnig F. 1986; Models for the structure of outer- membrane proteins of Escherichia coli derived from Raman spectroscopy and prediction methods. J Mol Biol 190:191–199
    [Google Scholar]
  53. Weller D.M., Cook R. J. 1983; Suppression of take-all of wheat by seed treatments with fluorescent Pseudomonads. Phytopathology 73:463–469
    [Google Scholar]
  54. Won J., Griffith R. W. 1993; Cloning and sequencing of the gene encoding a 31-kilodalton antigen of Haemophilus somnus. Infect Immun 61:2813–2821
    [Google Scholar]
  55. Wong R.S.Y., Jost H., Hancock R. E. W. 1993; Linker-insertion mutagenesis of Pseudomonas aeruginosa outer membrane protein Opr F. Mol Microbiol 10:283–292
    [Google Scholar]
  56. Wong W.C., Preece T. F. 1979; Identification of Pseudomonas tolaasir. the white line in agar and mushroom tissue block rapid pitting tests. J Appl Bacteriol 47:401–407
    [Google Scholar]
  57. Woodruff W.A., Hancock R. E. W. 1989; Pseudomonas aeruginosa outer membrane protein F: structural role and relationship to the Escherichia coli Omp A protein. J Bacteriol 171:3304–3309
    [Google Scholar]
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