Penicillin-binding Components Research Articles

Penicillin-binding components of penicillin-susceptible and -resistant strains of Streptococcus pneumoniae.

Three strains of Streptococcus pneumoniae with low-level resistance to penicillin, one strain with resistance to penicillin, and three strains susceptible to penicillin were compared. The three susceptible strains had very similar patterns of penicillin-binding components (PBCs) as detected by fluorography after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gels of these strains showed four major bands and a single minor band. All of the resistant strains showed different PBC patterns from those obtained for susceptible strains. The most marked changes were observed with the most resistant strain SPR110, which showed markedly reduced intensity of PBC 3 and a reduction in intensity of PBCs 1 and 2, as well as additional bands in the region of band 2. Low-level resistant strains had band densities different from the susceptible strains principally for PBC 3. No beta-lactamase activity or plasmid was detected in the resistant strains. An apparent difference in affinity for cloxacillin was also demonstrated between some of the PBCs of susceptible strain S. pneumoniae SPS101 and resistant strain S. pneumoniae SPR110. Penicillin resistance in the low-level resistant and the resistant strains was associated with PBCs possessing different properties from those seen in the three susceptible strains. Reduction in affinity for penicillin or a reduction in the amount of a PBC protein, or both, are the probable explanations of penicillin resistance in these strains.

Effect of β-Lactam Antibiotics on In Vitro Peptidoglycan Cross-Linking by a Particulate Fraction from Escherichia coli K-12 and Bacillus megaterium KM

The binding constants of several beta-lactam antibiotics towards penicillin-binding components in Escherichia coli K-12 (Spratt, Eur. J. Biochem. 72:341-352, 1977) and the antibiotic concentrations required to inhibit the peptidoglycan transpeptidase of E. coli 50% were compared. Penicillin-binding component 1B may have been the transpeptidase working in vitro. The structure-activity relationships of beta-lactam antibiotics and the mechanisms of action in E. coli and Bacillus megaterium are discussed.

On the process of cellular division in Escherichia coli: a series of mutants of E. coli altered in the penicillin-binding proteins.

A series of mutants defective in penicillin-binding components (or protein) (PBCs) was isolated from a collection of thermosensitive mutants of Escherichia coli. The mutants included mutations for each PBC (1 through 4) and a mutation in the activity of D-alanine carboxypeptidase 1a (PBC-5/6). PBC-1 was resolved into two components, PBC-1a and PBC-1b, which were the products of different genes referred to as ponA and ponB, respectively. No mutation examined in ponA-- or ponB--, by itself, was associated with a thermosensitive defect in growth. The mutants having ponB-- were hypersensitive to cephalosporins, which showed unusually high affinity to PBC-1a. Murein synthetic activity decreased markedly in cell-free preparations from the ponB-- mutant. The double mutation ponA-- ponB-- was found to be lethal. A mutant having ponAts and ponB-- revealed thermosensitivity of growth and lysed at the restrictive temperature. It is concluded that PBC-1a and PBC-1b could share a biochemical reaction necessary for cell elongation so that the function of either one may bypass the loss of the function of the other. With the series of mutants, the chromosomal locus of the gene for each PBC was mapped: ponA (PBC-1a) 73.5 min, ponB (PBC-1b) 3.3 min, rodA (PBC-2) 14.4 min, ftsI (PBC-3) 1.8 min, dacB (PBC-4) 68 min, and dacA (PBC-5/6) 13.7 min.

Studies on the penicillin-binding components of Bacillus megaterium

FEBS LettersVolume 76, Issue 2 p. 199-203 Full-length articleFree Access Studies on the penicillin-binding components of Bacillus megaterium Howard A. Chase, Howard A. Chase Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this authorStephen T. Shepherd, Stephen T. Shepherd Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this authorPeter E. Reynolds, Peter E. Reynolds Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this author Howard A. Chase, Howard A. Chase Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this authorStephen T. Shepherd, Stephen T. Shepherd Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this authorPeter E. Reynolds, Peter E. Reynolds Sub-department of Chemical Microbiology, Department of Biochemistry, Tennis Court Road, Cambridge, CB2 1QW, EnglandSearch for more papers by this author First published: April 15, 1977 https://doi.org/10.1016/0014-5793(77)80151-8Citations: 29AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume76, Issue2April 15, 1977Pages 199-203 ReferencesRelatedInformation

Mode of action of penicillins in vivo and in vitro in Bacillus megaterium.

A new system in which the in vivo and in vitro formation of cross-links in the peptidoglycan of Bacillus megaterium can be compared directly has been developed. The method for the determination of the in vivo cross-linking consists of lysozyme digestion of acetylated [(14)C]diaminopimelic acid-labeled cells and Bio-Gel P-6 gel filtration of the digest. The elution profile indicates the cell wall synthesized in vivo consists of highly cross-linked fractions (44%), bisdisaccharide peptide(s) (38%), and disaccharide peptide(s) (18%). The in vitro system showed a high synthetic activity of cross-linked peptidoglycan. The synthesis was inhibited completely by 83.3 mug of ristocetin or vancomycin per ml or 10(-4) M p-chloromercuribenzoate and inhibited only partially by penicillins. The polymerization was stimulated by high concentrations of sucrose, glycerol, amino acids, or dimethyl sulfoxide. The formation of cross-links was inhibited 50% at 0.3 mug of dicloxacillin per ml and 90% at 0.5 mug or more. It was also stimulated by high concentrations of sucrose, glycerol, or dimethyl sulfoxide. Effective concentrations of dicloxacillin on the growth, viability, and morphology of B. megaterium were determined. Sharp inhibition of cross-linking occurred in vivo and in vitro at these effective concentrations, whereas the incorporation of [(14)C]-diaminopimelate into bacterial cells was not affected at all. Cell-bound dicloxacillin reduced severely the degree of cross-linking in the cell wall synthesized after transfer to a dicloxacillin-free medium. Cell wall synthesized in the presence of dicloxacillin showed a higher rate of turnover than did the normal cell wall. Moreover, disaccharide peptide(s) was degraded faster than was bisdisaccharide peptide(s) in dicloxacillin-treated cells. From these observations, the primary target of penicillin action in B. megaterium is discussed in relation to the inhibition of cross-linking, penicillin-binding components, and cell lysis.

Altered penicillin-binding components in penicillin-resistant mutants of Bacillus subtilis.

Penicillin- (cloxacillin-) resistant mutants of Bacillus subtilis were isolated in a stepwise fashion and the five penicillin-binding components (PBCs) in each were examined to determine which of the proteins, if any, corresponds to the penicillin killing site. PBCs II and V were previously eliminated as the likely penicillin targett. In the present work, PBC IV showed no change in sensitivity to cloxacillin in any of the resistant mutants isolated. PBC I did not change until the fifth-step mutant, in which it could not be detected by penicillin binding. Since PBC I did not bind penicillins that are lethal for this mutant, it also cannot be the lethal target. PBC II showed increased resistance to cloxacillin in three discrete steps, i.e., in mutants 1, 4, and 5, accompanied by changes in its electrophoretic mobility. However, the sensitivity of PBC II to penicillin G changed very little. Correspondingly, the cloxacillin-resistant mutants were unaltered in their sensitivity to penicillin G in vivo. Thus, of the five PBCs found in B. subtilis, PBC II is the most likely target for killing by penicillins.

PENICILLIN‐SENSITIVE ENZYMES AND PENICILLIN‐BINDING COMPONENTS IN BACTERIAL CELLS*

Annals of the New York Academy of SciencesVolume 235, Issue 1 p. 210-224 PENICILLIN-SENSITIVE ENZYMES AND PENICILLIN-BINDING COMPONENTS IN BACTERIAL CELLS* Jack L. Strominger, Jack L. Strominger The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorEileen Willoughby, Eileen Willoughby The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorTatsuyuki Kamiryo, Tatsuyuki Kamiryo The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorPeter M. Blumberg, Peter M. Blumberg The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorR. Rogers Yocum, R. Rogers Yocum The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this author Jack L. Strominger, Jack L. Strominger The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorEileen Willoughby, Eileen Willoughby The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorTatsuyuki Kamiryo, Tatsuyuki Kamiryo The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorPeter M. Blumberg, Peter M. Blumberg The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this authorR. Rogers Yocum, R. Rogers Yocum The Biological Laboratories Harvard University Cambridge, Massachusetts 02138Search for more papers by this author First published: May 1974 https://doi.org/10.1111/j.1749-6632.1974.tb43267.xCitations: 41 * Work in our laboratory has been generously supported by Research Grants from the United States Public Health Service (AI-09152 and AM-13230) and from the National Science Foundation (GB-29747 and GB-30690). AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume235, Issue1Mode of Action of Antibiotics on Microbial Walls and MembranesMay 1974Pages 210-224 RelatedInformation

Penicillin binding components in Bacillus subtilis during sporulation.

The penicillin binding components of sporulating Bacillus subtilis 3610 were compared with those of the vegetative organism. No qualitative difference in binding components was observed. The total binding, however, varied with the phase of sporulation.

Isolation by Covalent Affinity Chromatography of the Penicillin-Binding Components from Membranes of Bacillus subtilis

An affinity chromatography technique was developed to isolate the five penicillin-binding components present in Bacillus subtilis membranes. The proteins were solubilized by the detergent Nonidet P-40, bound covalently to penicillin-substituted Sepharose, and subsequently eluted from the matrix with neutral hydroxylamine, which cleaves the penicilloyl-enzyme bond. Penicillin binding-component V, the D-alanine carboxypeptidase, makes up 1% of the total membrane protein. A modification of the above procedure enabled this enzyme to be obtained from the membrane in pure form in a single step with 50% overall recovery of enzymatic activity.

Five Penicillin-binding Components Occur in Bacillus subtilis Membranes

Abstract Membranes of Bacillus subtilis to which [14C]penicillin G had been bound were fractionated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Five labeled components were separated. These five components differed from each other in the rates at which they bound penicillins and cephalosporins by up to four orders of magnitude. The major penicillin-binding component, V, was identified as the d-alanine carboxypeptidase, whose activity is not vital for the cell. Three of the other components, in contrast, bound penicillins and cephalosporins at rates comparable to that of the penicillin killing site in this organism.

Multiple Penicillin-binding Components in Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Escherichia coli

Abstract A variety of techniques have been employed to investigate the penicillin-binding components of Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Escherichia coli. Virtually all the binding sites in these organisms are located in the cell membrane. All the techniques which have been employed suggest that more than one binding component exists in each of these organisms, although S. aureus and E. coli both appear to have only a single major one. A particularly interesting observation was that in Bacillus subtilis one of the two major components which reacts with penicillin G does not react readily with cephalothin.