Abstract
A recombinant plasmid containing ompK36, the gene coding for the Klebsiella pneumoniae outer membrane protein OmpK36, was constructed by transposon mutagenesis and subcloning. Clones were identified in a cosmid library in Escherichia coli on the basis of their reaction with antiserum against the OmpK36 protein and by the presence in gel electrophoretic analysis of a band in E. coli outer membranes migrating with a mobility corresponding to 36 kDa. The ompK36-encoded protein exhibited characteristic properties of porins, such as heat modifiability and resistance to trypsin. The sequence of the gene revealed that OmpK36 is a close relative of the enterobacterial porin family, with a high degree of homology with E. coli OmpC, PhoE, and OmpF. On the basis of the structures of OmpF and PhoE porins, determined previously by X-ray analysis, it appears likely that the three-dimensional structure of OmpK36 also contains the motif of a 16-stranded beta-barrel, with long loops on one end and short turns on the other. Like the OmpC porin from E. coli, OmpK36 contains a long insertion in loop 4. The results of a binding study of complement component C1q to OmpK36 and the analysis of the OmpK36 model suggest that C1q binding sites are covered by the lipopolysaccharide core in the native porin.
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- Agüero J., Mora G., Mroczenski-Wildey M. J., Fernandez-Beros M. E., Aron L., Cabello F. C. Cloning, expression and characterization of the 36 KDal Salmonella typhi porin gene in Escherichia coli. Microb Pathog. 1987 Dec;3(6):399–407. doi: 10.1016/0882-4010(87)90010-6. [DOI] [PubMed] [Google Scholar]
- Albertí S., Marqués G., Camprubí S., Merino S., Tomás J. M., Vivanco F., Benedí V. J. C1q binding and activation of the complement classical pathway by Klebsiella pneumoniae outer membrane proteins. Infect Immun. 1993 Mar;61(3):852–860. doi: 10.1128/iai.61.3.852-860.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
- Bodey G. P., Elting L. S., Rodriquez S., Hernandez M. Klebsiella bacteremia. A 10-year review in a cancer institution. Cancer. 1989 Dec 1;64(11):2368–2376. doi: 10.1002/1097-0142(19891201)64:11<2368::aid-cncr2820641129>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Click E. M., McDonald G. A., Schnaitman C. A. Translational control of exported proteins that results from OmpC porin overexpression. J Bacteriol. 1988 May;170(5):2005–2011. doi: 10.1128/jb.170.5.2005-2011.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cole S. T., Sonntag I., Henning U. Cloning and expression in Escherichia coli K-12 of the genes for major outer membrane protein OmpA from Shigella dysenteriae, Enterobacter aerogenes, and Serratia marcescens. J Bacteriol. 1982 Jan;149(1):145–150. doi: 10.1128/jb.149.1.145-150.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cowan S. W., Schirmer T., Rummel G., Steiert M., Ghosh R., Pauptit R. A., Jansonius J. N., Rosenbusch J. P. Crystal structures explain functional properties of two E. coli porins. Nature. 1992 Aug 27;358(6389):727–733. doi: 10.1038/358727a0. [DOI] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dhaese P., De Greve H., Decraemer H., Schell J., Van Montagu M. Rapid mapping of transposon insertion and deletion mutations in the large Ti-plasmids of Agrobacterium tumefaciens. Nucleic Acids Res. 1979 Dec 11;7(7):1837–1849. doi: 10.1093/nar/7.7.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duncan A. R., Winter G. The binding site for C1q on IgG. Nature. 1988 Apr 21;332(6166):738–740. doi: 10.1038/332738a0. [DOI] [PubMed] [Google Scholar]
- Figueroa J. E., Densen P. Infectious diseases associated with complement deficiencies. Clin Microbiol Rev. 1991 Jul;4(3):359–395. doi: 10.1128/cmr.4.3.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galdiero F., Tufano M. A., Sommese L., Folgore A., Tedesco F. Activation of complement system by porins extracted from Salmonella typhimurium. Infect Immun. 1984 Nov;46(2):559–563. doi: 10.1128/iai.46.2.559-563.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grant S. G., Jessee J., Bloom F. R., Hanahan D. Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4645–4649. doi: 10.1073/pnas.87.12.4645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hervás J. A., Alomar A., Salvá F., Reina J., Benedí V. J. Neonatal sepsis and meningitis in Mallorca, Spain, 1977-1991. Clin Infect Dis. 1993 May;16(5):719–724. doi: 10.1093/clind/16.5.719. [DOI] [PubMed] [Google Scholar]
- Hirsch P. R., Beringer J. E. A physical map of pPH1JI and pJB4JI. Plasmid. 1984 Sep;12(2):139–141. doi: 10.1016/0147-619x(84)90059-3. [DOI] [PubMed] [Google Scholar]
- Jeanteur D., Lakey J. H., Pattus F. The bacterial porin superfamily: sequence alignment and structure prediction. Mol Microbiol. 1991 Sep;5(9):2153–2164. doi: 10.1111/j.1365-2958.1991.tb02145.x. [DOI] [PubMed] [Google Scholar]
- Joiner K. A. Complement evasion by bacteria and parasites. Annu Rev Microbiol. 1988;42:201–230. doi: 10.1146/annurev.mi.42.100188.001221. [DOI] [PubMed] [Google Scholar]
- Kaneko M., Yamaguchi A., Sawai T. Purification and characterization of two kinds of porins from the Enterobacter cloacae outer membrane. J Bacteriol. 1984 Jun;158(3):1179–1181. doi: 10.1128/jb.158.3.1179-1181.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
- Loos M., Clas F. Antibody-independent killing of gram-negative bacteria via the classical pathway of complement. Immunol Lett. 1987 Feb;14(3):203–208. doi: 10.1016/0165-2478(87)90102-7. [DOI] [PubMed] [Google Scholar]
- Neidhardt F. C., Vaughn V., Phillips T. A., Bloch P. L. Gene-protein index of Escherichia coli K-12. Microbiol Rev. 1983 Jun;47(2):231–284. doi: 10.1128/mr.47.2.231-284.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nikaido H. Porins and specific diffusion channels in bacterial outer membranes. J Biol Chem. 1994 Feb 11;269(6):3905–3908. [PubMed] [Google Scholar]
- Nikaido H. Proteins forming large channels from bacterial and mitochondrial outer membranes: porins and phage lambda receptor protein. Methods Enzymol. 1983;97:85–100. doi: 10.1016/0076-6879(83)97122-7. [DOI] [PubMed] [Google Scholar]
- Nikaido H., Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rev. 1985 Mar;49(1):1–32. doi: 10.1128/mr.49.1.1-32.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pangon B., Bizet C., Buré A., Pichon F., Philippon A., Regnier B., Gutmann L. In vivo selection of a cephamycin-resistant, porin-deficient mutant of Klebsiella pneumoniae producing a TEM-3 beta-lactamase. J Infect Dis. 1989 May;159(5):1005–1006. doi: 10.1093/infdis/159.5.1005. [DOI] [PubMed] [Google Scholar]
- Puente J. L., Verdugo-Rodríguez A., Calva E. Expression of Salmonella typhi and Escherichia coli OmpC is influenced differently by medium osmolarity; dependence on Escherichia coli OmpR. Mol Microbiol. 1991 May;5(5):1205–1210. doi: 10.1111/j.1365-2958.1991.tb01894.x. [DOI] [PubMed] [Google Scholar]
- Rice L. B., Carias L. L., Etter L., Shlaes D. M. Resistance to cefoperazone-sulbactam in Klebsiella pneumoniae: evidence for enhanced resistance resulting from the coexistence of two different resistance mechanisms. Antimicrob Agents Chemother. 1993 May;37(5):1061–1064. doi: 10.1128/aac.37.5.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rietschel E. T., Kirikae T., Schade F. U., Ulmer A. J., Holst O., Brade H., Schmidt G., Mamat U., Grimmecke H. D., Kusumoto S. The chemical structure of bacterial endotoxin in relation to bioactivity. Immunobiology. 1993 Apr;187(3-5):169–190. doi: 10.1016/S0171-2985(11)80338-4. [DOI] [PubMed] [Google Scholar]
- Stemmer F., Loos M. Evidence for direct binding of the first component of complement, C1, to outer membrane proteins from Salmonella minnesota. Curr Top Microbiol Immunol. 1985;121:73–84. doi: 10.1007/978-3-642-45604-6_4. [DOI] [PubMed] [Google Scholar]
- Tomás J. M., Benedí V. J., Ciurana B., Jofre J. Role of capsule and O antigen in resistance of Klebsiella pneumoniae to serum bactericidal activity. Infect Immun. 1986 Oct;54(1):85–89. doi: 10.1128/iai.54.1.85-89.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tomás J. M., Jofre J. T. Lipopolysaccharide-specific bacteriophage for Klebsiella pneumoniae C3. J Bacteriol. 1985 Jun;162(3):1276–1279. doi: 10.1128/jb.162.3.1276-1279.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van der Ley P., Bekkers A., Van Meersbergen J., Tommassen J. A comparative study on the phoE genes of three enterobacterial species. Implications for structure-function relationships in a pore-forming protein of the outer membrane. Eur J Biochem. 1987 Apr 15;164(2):469–475. doi: 10.1111/j.1432-1033.1987.tb11080.x. [DOI] [PubMed] [Google Scholar]
- Werts C., Charbit A., Bachellier S., Hofnung M. DNA sequence analysis of the lamB gene from Klebsiella pneumoniae: implications for the topology and the pore functions in maltoporin. Mol Gen Genet. 1992 Jun;233(3):372–378. doi: 10.1007/BF00265433. [DOI] [PubMed] [Google Scholar]
- Woodruff W. A., Parr T. R., Jr, Hancock R. E., Hanne L. F., Nicas T. I., Iglewski B. H. Expression in Escherichia coli and function of Pseudomonas aeruginosa outer membrane porin protein F. J Bacteriol. 1986 Aug;167(2):473–479. doi: 10.1128/jb.167.2.473-479.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Bruijn F. J., Lupski J. R. The use of transposon Tn5 mutagenesis in the rapid generation of correlated physical and genetic maps of DNA segments cloned into multicopy plasmids--a review. Gene. 1984 Feb;27(2):131–149. doi: 10.1016/0378-1119(84)90135-5. [DOI] [PubMed] [Google Scholar]
- van de Klundert J. A., van Gestel M. H., Meerdink G., de Marie S. Emergence of bacterial resistance to cefamandole in vivo due to outer membrane protein deficiency. Eur J Clin Microbiol Infect Dis. 1988 Dec;7(6):776–778. doi: 10.1007/BF01975046. [DOI] [PubMed] [Google Scholar]