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. 1994 Jan;176(2):359–367. doi: 10.1128/jb.176.2.359-367.1994

New outer membrane-associated protease of Escherichia coli K-12.

A Kaufmann 1, Y D Stierhof 1, U Henning 1
PMCID: PMC205058  PMID: 8288530

Abstract

The gene for a new outer membrane-associated protease, designated OmpP, of Escherichia coli has been cloned and sequenced. The gene encodes a 315-residue precursor protein possessing a 23-residue signal sequence. Including conservative substitutions and omitting the signal peptides, OmpP is 87% identical to the outer membrane protease OmpT. OmpP possessed the same enzymatic activity as OmpT. Immuno-electron microscopy demonstrated the exposure of the protein at the cell surface. Digestion of intact cells with proteinase K removed 155 N-terminal residues of OmpP, while the C-terminal half remained protected. It is possible that much of this N-terminal part is cell surface exposed and carries the enzymatic activity. Synthesis of OmpP was found to be thermoregulated, as is the expression of ompT (i.e., there is a low rate of synthesis at low temperatures) and, in addition, was found to be controlled by the cyclic AMP system.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  2. 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]
  3. Chen R., Schmidmayr W., Krämer C., Chen-Schmeisser U., Henning U. Primary structure of major outer membrane protein II (ompA protein) of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4592–4596. doi: 10.1073/pnas.77.8.4592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Drexler K., Dannull J., Hindennach I., Mutschler B., Henning U. Single mutations in a gene for a tail fiber component of an Escherichia coli phage can cause an extension from a protein to a carbohydrate as a receptor. J Mol Biol. 1991 Jun 20;219(4):655–663. doi: 10.1016/0022-2836(91)90662-p. [DOI] [PubMed] [Google Scholar]
  7. Dumont M. E., Trewhella J., Engelman D. M., Richards F. M. Stability of transmembrane regions in bacteriorhodopsin studied by progressive proteolysis. J Membr Biol. 1985;88(3):233–247. doi: 10.1007/BF01871088. [DOI] [PubMed] [Google Scholar]
  8. Freudl R., Schwarz H., Stierhof Y. D., Gamon K., Hindennach I., Henning U. An outer membrane protein (OmpA) of Escherichia coli K-12 undergoes a conformational change during export. J Biol Chem. 1986 Aug 25;261(24):11355–11361. [PubMed] [Google Scholar]
  9. Georgellis D., Arvidson S., von Gabain A. Decay of ompA mRNA and processing of 9S RNA are immediately affected by shifts in growth rate, but in opposite manners. J Bacteriol. 1992 Aug;174(16):5382–5390. doi: 10.1128/jb.174.16.5382-5390.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grodberg J., Dunn J. J. Comparison of Escherichia coli K-12 outer membrane protease OmpT and Salmonella typhimurium E protein. J Bacteriol. 1989 May;171(5):2903–2905. doi: 10.1128/jb.171.5.2903-2905.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grodberg J., Dunn J. J. ompT encodes the Escherichia coli outer membrane protease that cleaves T7 RNA polymerase during purification. J Bacteriol. 1988 Mar;170(3):1245–1253. doi: 10.1128/jb.170.3.1245-1253.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grodberg J., Lundrigan M. D., Toledo D. L., Mangel W. F., Dunn J. J. Complete nucleotide sequence and deduced amino acid sequence of the ompT gene of Escherichia coli K-12. Nucleic Acids Res. 1988 Feb 11;16(3):1209–1209. doi: 10.1093/nar/16.3.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gunasekera A., Ebright Y. W., Ebright R. H. DNA sequence determinants for binding of the Escherichia coli catabolite gene activator protein. J Biol Chem. 1992 Jul 25;267(21):14713–14720. [PubMed] [Google Scholar]
  14. Hancock R. E., Reeves P. Bacteriophage resistance in Escherichia coli K-12: general pattern of resistance. J Bacteriol. 1975 Mar;121(3):983–993. doi: 10.1128/jb.121.3.983-993.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  16. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hollifield W. C., Jr, Fiss E. H., Neilands J. B. Modification of a ferric enterobactin receptor protein from the outer membrane of Escherichia coli. Biochem Biophys Res Commun. 1978 Jul 28;83(2):739–746. doi: 10.1016/0006-291x(78)91051-3. [DOI] [PubMed] [Google Scholar]
  18. Jennings M. L. Topography of membrane proteins. Annu Rev Biochem. 1989;58:999–1027. doi: 10.1146/annurev.bi.58.070189.005031. [DOI] [PubMed] [Google Scholar]
  19. KAY D., FILDES P. Hydroxymethylcytosine-containing and tryptophan-dependent bacteriophages isolated from city effluents. J Gen Microbiol. 1962 Jan;27:143–146. doi: 10.1099/00221287-27-1-143. [DOI] [PubMed] [Google Scholar]
  20. Klauser T., Pohlner J., Meyer T. F. Extracellular transport of cholera toxin B subunit using Neisseria IgA protease beta-domain: conformation-dependent outer membrane translocation. EMBO J. 1990 Jun;9(6):1991–1999. doi: 10.1002/j.1460-2075.1990.tb08327.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klauser T., Pohlner J., Meyer T. F. Selective extracellular release of cholera toxin B subunit by Escherichia coli: dissection of Neisseria Iga beta-mediated outer membrane transport. EMBO J. 1992 Jun;11(6):2327–2335. doi: 10.1002/j.1460-2075.1992.tb05292.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klose M., Schwarz H., MacIntyre S., Freudl R., Eschbach M. L., Henning U. 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. 1988 Sep 15;263(26):13291–13296. [PubMed] [Google Scholar]
  23. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lederberg E M, Lederberg J. Genetic Studies of Lysogenicity in Escherichia Coli. Genetics. 1953 Jan;38(1):51–64. doi: 10.1093/genetics/38.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leong J., Neilands J. B. Mechanisms of siderophore iron transport in enteric bacteria. J Bacteriol. 1976 May;126(2):823–830. doi: 10.1128/jb.126.2.823-830.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. MacIntyre S., Mutschler B., Henning U. Requirement of the SecB chaperone for export of a non-secretory polypeptide in Escherichia coli. Mol Gen Genet. 1991 Jun;227(2):224–228. doi: 10.1007/BF00259674. [DOI] [PubMed] [Google Scholar]
  28. Morona R., Henning U. Host range mutants of bacteriophage Ox2 can use two different outer membrane proteins of Escherichia coli K-12 as receptors. J Bacteriol. 1984 Aug;159(2):579–582. doi: 10.1128/jb.159.2.579-582.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Morona R., Klose M., Henning U. Escherichia coli K-12 outer membrane protein (OmpA) as a bacteriophage receptor: analysis of mutant genes expressing altered proteins. J Bacteriol. 1984 Aug;159(2):570–578. doi: 10.1128/jb.159.2.570-578.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Morona R., Tommassen J., Henning U. Demonstration of a bacteriophage receptor site on the Escherichia coli K12 outer-membrane protein OmpC by the use of a protease. Eur J Biochem. 1985 Jul 1;150(1):161–169. doi: 10.1111/j.1432-1033.1985.tb09002.x. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Pacaud M. Identification and localization of two membrane-bound esterases from Escherichia coli. J Bacteriol. 1982 Jan;149(1):6–14. doi: 10.1128/jb.149.1.6-14.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pacaud M. Purification and characterization of two novel proteolytic enzymes in membranes of Escherichia coli. Protease IV and protease V. J Biol Chem. 1982 Apr 25;257(8):4333–4339. [PubMed] [Google Scholar]
  34. Palmer S. M., St John A. C. Characterization of a membrane-associated serine protease in Escherichia coli. J Bacteriol. 1987 Apr;169(4):1474–1479. doi: 10.1128/jb.169.4.1474-1479.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pohlner J., Halter R., Beyreuther K., Meyer T. F. Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease. 1987 Jan 29-Feb 4Nature. 325(6103):458–462. doi: 10.1038/325458a0. [DOI] [PubMed] [Google Scholar]
  36. Pugsley A. P., d'Enfert C., Reyss I., Kornacker M. G. Genetics of extracellular protein secretion by gram-negative bacteria. Annu Rev Genet. 1990;24:67–90. doi: 10.1146/annurev.ge.24.120190.000435. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Rupprecht K. R., Gordon G., Lundrigan M., Gayda R. C., Markovitz A., Earhart C. omp T: Escherichia coli K-12 structural gene for protein a (3b). J Bacteriol. 1983 Feb;153(2):1104–1106. doi: 10.1128/jb.153.2.1104-1106.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  40. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schnaitman C. A., McDonald G. A. Regulation of outer membrane protein synthesis in Escherichia coli K-12: deletion of ompC affects expression of the OmpF protein. J Bacteriol. 1984 Aug;159(2):555–563. doi: 10.1128/jb.159.2.555-563.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schwarz H., Riede I., Sonntag I., Henning U. Degrees of relatedness of T-even type E. coli phages using different or the same receptors and topology of serologically cross-reacting sites. EMBO J. 1983;2(3):375–380. doi: 10.1002/j.1460-2075.1983.tb01433.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Skurray R. A., Hancock R. E., Reeves P. Con--mutants: class of mutants in Escherichia coli K-12 lacking a major cell wall protein and defective in conjugation and adsorption of a bacteriophage. J Bacteriol. 1974 Sep;119(3):726–735. doi: 10.1128/jb.119.3.726-735.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Slot J. W., Geuze H. J. A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol. 1985 Jul;38(1):87–93. [PubMed] [Google Scholar]
  45. Sodeinde O. A., Goguen J. D. Nucleotide sequence of the plasminogen activator gene of Yersinia pestis: relationship to ompT of Escherichia coli and gene E of Salmonella typhimurium. Infect Immun. 1989 May;57(5):1517–1523. doi: 10.1128/iai.57.5.1517-1523.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sugimura K., Nishihara T. Purification, characterization, and primary structure of Escherichia coli protease VII with specificity for paired basic residues: identity of protease VII and OmpT. J Bacteriol. 1988 Dec;170(12):5625–5632. doi: 10.1128/jb.170.12.5625-5632.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vogel H., Jähnig F. Models for the structure of outer-membrane proteins of Escherichia coli derived from raman spectroscopy and prediction methods. J Mol Biol. 1986 Jul 20;190(2):191–199. doi: 10.1016/0022-2836(86)90292-5. [DOI] [PubMed] [Google Scholar]
  48. Weiss M. S., Kreusch A., Schiltz E., Nestel U., Welte W., Weckesser J., Schulz G. E. The structure of porin from Rhodobacter capsulatus at 1.8 A resolution. FEBS Lett. 1991 Mar 25;280(2):379–382. doi: 10.1016/0014-5793(91)80336-2. [DOI] [PubMed] [Google Scholar]
  49. Weiss M. S., Schulz G. E. Structure of porin refined at 1.8 A resolution. J Mol Biol. 1992 Sep 20;227(2):493–509. doi: 10.1016/0022-2836(92)90903-w. [DOI] [PubMed] [Google Scholar]
  50. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]
  51. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  52. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]
  54. Yu G. Q., Hong J. S. Identification and nucleotide sequence of the activator gene of the externally induced phosphoglycerate transport system of Salmonella typhimurium. Gene. 1986;45(1):51–57. doi: 10.1016/0378-1119(86)90131-9. [DOI] [PubMed] [Google Scholar]
  55. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

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