Improved high-performance liquid chromatographic separation of peptidoglycan isolated from various Staphylococcus aureus strains for mass spectrometric characterization

Roos, Martin; Pittenauer, Ernst; Schmid, E.; Beyer, M.; Reinike, Bernhard; Allmaier, Günter; Labischinski, Harald

doi:10.1016/s0378-4347(97)00506-9

Cited by 21 publications

(33 citation statements)

References 17 publications

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“…We found that the HPLC chromatograms for all strains, irrespective of resistance phenotype, had the typical muropeptide species identified in previous investigations (7,14,17). The nomenclature used to identify relevant peaks was described previously (17,22).…”

Section: Analysis Of Peptidoglycanssupporting

confidence: 60%

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A Spectrum of Changes Occurs in Peptidoglycan Composition of Glycopeptide-Intermediate Clinical Staphylococcus aureus Isolates

Boyle-Vavra

Labischinski

Ebert

et al. 2001

Antimicrob Agents Chemother

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The mechanism of glycopeptide resistance in Staphylococcus aureus is not known with certainty. Because the target of vancomycin is the D-Ala-D-Ala terminus of the stem peptide of the peptidoglycan precursor, by subjecting muropeptides to reversed-phase high-performance liquid chromatography, we investigated peptidoglycan obtained from glycopeptide-intermediate S. aureus (GISA) isolates for changes in composition and evaluated whether any peptidoglycan structural change was a consistent feature of clinical GISA isolates. GISA isolates Mu50 and Mu3 from Japan had the large glutamate-containing monomeric peak demonstrated previously, although strain H1, a vancomycin-susceptible MRSA isolate from Japan that was clonally related to Mu3 and Mu50, and a femC mutant that we studied, did also. For the U.S. GISA isolates, strain NJ had a large monomeric peak with a retention time identical to that described for the glutamate-containing monomer in strains H1, Mu3, and Mu50. However, a much smaller corresponding peak was seen in GISA MI, and this peak was absent from both GISA PC and a recent GISA isolate obtained from an adult patient in Illinois (strain IL). These data suggest that a uniform alteration in peptidoglycan composition cannot be discerned among the GISA isolates and indicate that a single genetic or biochemical change is unlikely to account for the glycopeptide resistance phenotype in the clinical GISA isolates observed to date. Furthermore, a large monomeric glutamate-containing peak is not sufficient to confer the resistance phenotype.

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Section: Analysis Of Peptidoglycanssupporting

confidence: 60%

“…The nomenclature used to identify relevant peaks was described previously (17,22). The dimeric, trimeric, etc., muropeptides represent peptide-cross-linked combinations of muropeptide monomeric structures.…”

Section: Analysis Of Peptidoglycansmentioning

confidence: 99%

A Spectrum of Changes Occurs in Peptidoglycan Composition of Glycopeptide-Intermediate Clinical Staphylococcus aureus Isolates

Boyle-Vavra

Labischinski

Ebert

et al. 2001

Antimicrob Agents Chemother

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“…Peptidoglycan preparations from the various strains were digested with muramidase, which cleaves after N-acetylmuramic acid residues, leaving peptide cross-bridges intact in the various disaccharide peptide units generated through the activity of the enzyme. The muropeptides generated were analyzed by HPLC, and peaks were identified and designated using previously described nomenclature (44,50). The proportions of muropeptide monomers, dimers, trimers, and oligomers are summarized in Table 1.…”

Section: Resultsmentioning

confidence: 99%

Cell Wall Composition and Decreased Autolytic Activity and Lysostaphin Susceptibility of Glycopeptide-Intermediate Staphylococcus aureus

Koehl

Muthaiyan

Jayaswal

et al. 2004

Antimicrob Agents Chemother

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The cell wall composition and autolytic properties of passage-selected glycopeptide-intermediate Staphylococcus aureus (GISA) isolates and their parent strains were studied in order to investigate the mechanism of decreased vancomycin susceptibility. GISA had relatively modest changes in peptidoglycan composition involving peptidoglycan interpeptide bridges and somewhat decreased cross-linking compared to that of parent strains. The cell wall phosphorus content of GISA strains was lower than that of susceptible parent strains, indicating somewhat lower wall teichoic acid levels in the GISA strains. Similar to whole cells, isolated crude cell walls retaining autolytic activity of GISA had drastically reduced autolytic activity compared to that of parent strains, and this arose early in the development of the GISA phenotype. This was due to an alteration in the autolytic enzymes of GISA as revealed by normal susceptibility of GISA-purified cell walls to parental strain autolysin extract and lower activity and altered peptidoglycan hydrolase activity profiles in GISA autolysin extracts compared to those of parent strains. Northern blot analysis indicated that expression of atl, the major autolysin gene, was significantly downregulated in a GISA strain compared to that of its parent strain. In contrast to whole cells, which showed decreased lysostaphin susceptibility, purified cell walls of GISA showed increased susceptibility to lysostaphin. We suggest that in our GISA strains, decreased autolytic activity is involved in the tolerance of vancomycin and the activities of endogenous autolysins are important in conferring sensitivity to lysostaphin on whole cells.

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“…2). HF treatment removes teichoic acids covalently bound to peptidoglycan (12). In S. pneumoniae, teichoic acids were reported to affect the bacteriolytic activity of pneumococcal autolysin (3,5).…”

Section: Resultsmentioning

confidence: 99%

Zymographic Characterization of Bacteriolytic Enzymes Produced by Oral Streptococci

Yoshimura

Komatsuzawa

Kajimura

et al. 2004

Microbiology and Immunology

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Zymographic analysis was performed to know the bacteriolytic enzyme profiles of 4% SDS extracts of oral streptococci, Streptococcus mutans, S. sobrinus, S. sanguis, S. mitis and S. salivarius. We investigated the five strains in each species and found that the profile was very similar among strains of the same species except for S. salivarius (the profile was classified into two types). On the other hand, the profile was considerably different among species. Two major bacteriolytic enzymes of S. mutans showing molecular mass of 80 and 100 kDa were found using SDS‐boiled S. mutans or S. sobrinus cells as substrate. These bacteriolytic activities were less apparent in the gel containing S. mitis or S. salivarius, and also not detectable in the gel containing S. sanguis. S. sobrinus extract showed only one bacteriolytic band (78 kDa) as strong activity using S. sobrinus cells as substrate. S. sanguis extract showed no bacteriolytic bands using any streptococcal cells. Extracts of either S. mitis or S. salivarius showed weak activity by using respective strains as substrate.

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Improved high-performance liquid chromatographic separation of peptidoglycan isolated from various Staphylococcus aureus strains for mass spectrometric characterization

Cited by 21 publications

References 17 publications

A Spectrum of Changes Occurs in Peptidoglycan Composition of Glycopeptide-Intermediate Clinical Staphylococcus aureus Isolates

A Spectrum of Changes Occurs in Peptidoglycan Composition of Glycopeptide-Intermediate Clinical Staphylococcus aureus Isolates

Cell Wall Composition and Decreased Autolytic Activity and Lysostaphin Susceptibility of Glycopeptide-Intermediate Staphylococcus aureus

Zymographic Characterization of Bacteriolytic Enzymes Produced by Oral Streptococci

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