Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Jan;176(1):256–259. doi: 10.1128/jb.176.1.256-259.1994

Artifactual processing of penicillin-binding proteins 7 and 1b by the OmpT protease of Escherichia coli.

T A Henderson 1, P M Dombrosky 1, K D Young 1
PMCID: PMC205039  PMID: 8282705

Abstract

Penicillin-binding proteins (PBPs) were visualized in strains of Escherichia coli that carried mutations in one or more of the following protease genes: tsp, degP, ptr, and ompT. In the absence of a functional ompT gene, PBPs 1b alpha and 7 were not processed to the shortened forms 1b beta and 8, respectively. Cleavage of PBPs 1b alpha and 7 could be restored by introduction of a plasmid carrying the wild-type ompT gene. These PBPs were processed only after cell lysis or after membrane perturbation of whole cells by freeze-thaw, suggesting that the cleavage was a nonspecific artifact due to contact with OmpT, an outer membrane protease, and that such processing was not biologically significant in vivo. The degradation of other PBPs during purification or storage may also be effected by OmpT.

Full text

PDF
256

Images in this article

257Image
on p.257
258Image
on p.258
258Image
on p.258

Selected References

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

  1. Akiyama Y., Ito K. SecY protein, a membrane-embedded secretion factor of E. coli, is cleaved by the ompT protease in vitro. Biochem Biophys Res Commun. 1990 Mar 16;167(2):711–715. doi: 10.1016/0006-291x(90)92083-c. [DOI] [PubMed] [Google Scholar]
  2. Baneyx F., Georgiou G. Construction and characterization of Escherichia coli strains deficient in multiple secreted proteases: protease III degrades high-molecular-weight substrates in vivo. J Bacteriol. 1991 Apr;173(8):2696–2703. doi: 10.1128/jb.173.8.2696-2703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baneyx F., Georgiou G. In vivo degradation of secreted fusion proteins by the Escherichia coli outer membrane protease OmpT. J Bacteriol. 1990 Jan;172(1):491–494. doi: 10.1128/jb.172.1.491-494.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barbas J. A., Díaz J., Rodríguez-Tébar A., Vázquez D. Specific location of penicillin-binding proteins within the cell envelope of Escherichia coli. J Bacteriol. 1986 Jan;165(1):269–275. doi: 10.1128/jb.165.1.269-275.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frère J. M., Joris B. Penicillin-sensitive enzymes in peptidoglycan biosynthesis. Crit Rev Microbiol. 1985;11(4):299–396. doi: 10.3109/10408418409105906. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hara H., Nishimura Y., Kato J., Suzuki H., Nagasawa H., Suzuki A., Hirota Y. Genetic analyses of processing involving C-terminal cleavage in penicillin-binding protein 3 of Escherichia coli. J Bacteriol. 1989 Nov;171(11):5882–5889. doi: 10.1128/jb.171.11.5882-5889.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kato J., Suzuki H., Hirota Y. Overlapping of the coding regions for alpha and gamma components of penicillin-binding protein 1 b in Escherichia coli. Mol Gen Genet. 1984;196(3):449–457. doi: 10.1007/BF00436192. [DOI] [PubMed] [Google Scholar]
  9. Leidenix M. J., Jacoby G. H., Henderson T. A., Young K. D. Separation of Escherichia coli penicillin-binding proteins into different membrane vesicles by agarose electrophoresis and sizing chromatography. J Bacteriol. 1989 Oct;171(10):5680–5686. doi: 10.1128/jb.171.10.5680-5686.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Masson J. M., Labia R. Synthesis of a 125I-radiolabeled penicillin for penicillin-binding proteins studies. Anal Biochem. 1983 Jan;128(1):164–168. doi: 10.1016/0003-2697(83)90357-3. [DOI] [PubMed] [Google Scholar]
  11. Mottl H., Terpstra P., Keck W. Penicillin-binding protein 4 of Escherichia coli shows a novel type of primary structure among penicillin-interacting proteins. FEMS Microbiol Lett. 1991 Mar 1;62(2-3):213–220. doi: 10.1016/0378-1097(91)90160-c. [DOI] [PubMed] [Google Scholar]
  12. Nagasawa H., Sakagami Y., Suzuki A., Suzuki H., Hara H., Hirota Y. Determination of the cleavage site involved in C-terminal processing of penicillin-binding protein 3 of Escherichia coli. J Bacteriol. 1989 Nov;171(11):5890–5893. doi: 10.1128/jb.171.11.5890-5893.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nakamura M., Maruyama I. N., Soma M., Kato J., Suzuki H., Horota Y. On the process of cellular division in Escherichia coli: nucleotide sequence of the gene for penicillin-binding protein 3. Mol Gen Genet. 1983;191(1):1–9. doi: 10.1007/BF00330881. [DOI] [PubMed] [Google Scholar]
  14. Nicholas R. A., Strominger J. L., Suzuki H., Hirota Y. Identification of the active site in penicillin-binding protein 3 of Escherichia coli. J Bacteriol. 1985 Oct;164(1):456–460. doi: 10.1128/jb.164.1.456-460.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Silber K. R., Keiler K. C., Sauer R. T. Tsp: a tail-specific protease that selectively degrades proteins with nonpolar C termini. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):295–299. doi: 10.1073/pnas.89.1.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  17. Strauch K. L., Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1576–1580. doi: 10.1073/pnas.85.5.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Suzuki H., Kato J., Sakagami Y., Mori M., Suzuki A., Hirota Y. Conversion of the alpha component of penicillin-binding protein 1b to the beta component in Escherichia coli. J Bacteriol. 1987 Feb;169(2):891–893. doi: 10.1128/jb.169.2.891-893.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tuomanen E., Schwartz J. Penicillin-binding protein 7 and its relationship to lysis of nongrowing Escherichia coli. J Bacteriol. 1987 Nov;169(11):4912–4915. doi: 10.1128/jb.169.11.4912-4915.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yousif S. Y., Broome-Smith J. K., Spratt B. G. Lysis of Escherichia coli by beta-lactam antibiotics: deletion analysis of the role of penicillin-binding proteins 1A and 1B. J Gen Microbiol. 1985 Oct;131(10):2839–2845. doi: 10.1099/00221287-131-10-2839. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES