Abstract
The penicillin-binding proteins (PBPs) of Neisseria gonorrhoeae were investigated by using [3H]benzylpenicillin of high specific activity. This made it possible to label the PBPs both in cytoplasmic membranes and in the membranes of actively growing cells (in vivo labeling). A total of 20 strains isolated from different geographic locales showed the same pattern of three major PBPs, which had molecular weights of approximately 90,000, 63,000, and 48,000. Five clinical isolates of intrinsically penicillin-resistant gonococci each exhibited reduced penicillin binding of PBPs 1 and 2. The construction of an isogenic set of transformants with increasing levels of penicillin resistance indicated that the penA gene was associated with a decrease in penicillin binding fo PBP 2. Decreased binding to PBP 1 is likely to accompany the newly reported pem and tem genes, which govern to highest level of penicillin resistance.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Buchanan C. E., Strominger J. L. Altered penicillin-binding components in penicillin-resistant mutants of Bacillus subtilis. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1816–1820. doi: 10.1073/pnas.73.6.1816. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Curtis N. A., Orr D., Ross G. W., Boulton M. G. Competition of beta-lactam antibiotics for the penicillin-binding proteins of Pseudomonas aeruginosa, Enterobacter cloacae, Klebsiella aerogenes, Proteus rettgeri, and Escherichia coli: comparison with antibacterial activity and effects upon bacterial morphology. Antimicrob Agents Chemother. 1979 Sep;16(3):325–328. doi: 10.1128/aac.16.3.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dougherty T. J., Asmus A., Tomasz A. Specificity of DNA uptake in genetic transformation of gonococci. Biochem Biophys Res Commun. 1979 Jan 15;86(1):97–104. doi: 10.1016/0006-291x(79)90386-3. [DOI] [PubMed] [Google Scholar]
- Guymon L. F., Walstad D. L., Sparling P. F. Cell envelope alterations in antibiotic-sensitive and-resistant strains of Neisseria gonorrhoeae. J Bacteriol. 1978 Oct;136(1):391–401. doi: 10.1128/jb.136.1.391-401.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hakenbeck R., Tarpay M., Tomasz A. Multiple changes of penicillin-binding proteins in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Antimicrob Agents Chemother. 1980 Mar;17(3):364–371. doi: 10.1128/aac.17.3.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kozarich J. W., Nishino T., Willoughby E., Strominger J. L. Hydroxylaminolysis of penicillin binding componenets is enzymatically catalyzed. J Biol Chem. 1977 Nov 10;252(21):7525–7529. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- 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]
- Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
- Leive L. The barrier function of the gram-negative envelope. Ann N Y Acad Sci. 1974 May 10;235(0):109–129. doi: 10.1111/j.1749-6632.1974.tb43261.x. [DOI] [PubMed] [Google Scholar]
- Maier T. W., Zubrzycki L., Coyle M. B. Genetic analysis of drug resistance in Neisseria gonorrhoeae: identification and linkage relationships of loci controlling drug resistance. Antimicrob Agents Chemother. 1975 May;7(5):676–681. doi: 10.1128/aac.7.5.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noguchi H., Matsuhashi M., Mitsuhashi S. Comparative studies of penicillin-binding proteins in Pseudomonas aeruginosa and Escherichia coli. Eur J Biochem. 1979 Oct;100(1):41–49. doi: 10.1111/j.1432-1033.1979.tb02031.x. [DOI] [PubMed] [Google Scholar]
- Nolan R. D., Hildebrandt J. F. Comparison of the penicillin-binding proteins of different strains of Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1979 Sep;16(3):336–340. doi: 10.1128/aac.16.3.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Rodriguez W. J., Saz A. K. Differential binding of penicillin by membrane fractions from penicillin-susceptible and -resistant gonococci. Antimicrob Agents Chemother. 1978 Apr;13(4):589–597. doi: 10.1128/aac.13.4.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodriguez W., Saz A. K. Possible mechanism of decreased susceptibility of Neisseria gonorrhoeae to penicillin. Antimicrob Agents Chemother. 1975 Jun;7(6):788–792. doi: 10.1128/aac.7.6.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenthal R. S., Wright R. M., Sinha R. K. Extent of peptide cross-linking in the peptidoglycan of Neisseria gonorrhoeae. Infect Immun. 1980 Jun;28(3):867–875. doi: 10.1128/iai.28.3.867-875.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scudamore R. A., Beveridge T. J., Goldner M. Outer-membrane penetration barriers as components of intrinsic resistance to beta-lactam and other antibiotics in Escherichia coli K-12. Antimicrob Agents Chemother. 1979 Feb;15(2):182–189. doi: 10.1128/aac.15.2.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scudamore R. A., Beveridge T. J., Goldner M. Penetrability of the outer membrane of Neisseria gonorrhoeae in relation to acquired resistance to penicillin and other antibiotics. Antimicrob Agents Chemother. 1979 Jun;15(6):820–827. doi: 10.1128/aac.15.6.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spratt B. G. Escherichia coli resistance to beta-lactam antibiotics through a decrease in the affinity of a target for lethality. Nature. 1978 Aug 17;274(5672):713–715. doi: 10.1038/274713a0. [DOI] [PubMed] [Google Scholar]
- Spratt B. G. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. doi: 10.1111/j.1432-1033.1977.tb11258.x. [DOI] [PubMed] [Google Scholar]
- Tamura T., Imae Y., Strominger J. L. Purification to homogeneity and properties of two D-alanine carboxypeptidases I From Escherichia coli. J Biol Chem. 1976 Jan 25;251(2):414–423. [PubMed] [Google Scholar]
- Tipper D. J. Mode of action of beta-lactam antibiotics. Rev Infect Dis. 1979 Jan-Feb;1(1):39–54. doi: 10.1093/clinids/1.1.39. [DOI] [PubMed] [Google Scholar]
- Walstad D. L., Guymon L. F., Sparling P. F. Altered outer membrane protein in different colonial types of Neisseria gonorrhoeae. J Bacteriol. 1977 Mar;129(3):1623–1627. doi: 10.1128/jb.129.3.1623-1627.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warner P. F., Zubrzycki L. J., Chila M. Polygenes and modifier genes for tetracycline and penicillin resistance in Neisseria gonorrhoeae. J Gen Microbiol. 1980 Mar;117(1):103–110. doi: 10.1099/00221287-117-1-103. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- Wolf-Watz H., Elmros T., Normark S., Bloom G. D. Cell envelope of Neisseria gonorrhoeae: outer membrane and peptidoglycan composition of penicillin-sensitive and-resistant strains. Infect Immun. 1975 Jun;11(6):1332–1341. doi: 10.1128/iai.11.6.1332-1341.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zighelboim S., Tomasz A. Penicillin-binding proteins of multiply antibiotic-resistant South African strains of Streptococcus pneumoniae. Antimicrob Agents Chemother. 1980 Mar;17(3):434–442. doi: 10.1128/aac.17.3.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zimmermann W. Penetration of beta-lactam antibiotics into their target enzymes in Pseudomonas aeruginosa: comparison of a highly sensitive mutant with its parent strain. Antimicrob Agents Chemother. 1980 Jul;18(1):94–100. doi: 10.1128/aac.18.1.94. [DOI] [PMC free article] [PubMed] [Google Scholar]