Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1975 Oct;124(1):410–418. doi: 10.1128/jb.124.1.410-418.1975

Characterization of the presumed peptide cross-links in the soluble peptidoglycan fragments synthesized by protoplasts of Streptococcus faecalis.

R S Rosenthal, G D Shockman
PMCID: PMC235909  PMID: 809418

Abstract

Protoplasts of Streptococcus faecalis ATCC 9790 were produced with the aid of lysozyme, and the ability of these bodies to synthesize soluble, peptide cross-linked peptidoglycan (PG) fragments was examined. Lysozyme digests of PG isolated using gel filtration from the supernatant medium of protoplasts grown in the presence of [14C]acetate and L-[3H]lysine contained small amounts of PG having KD expected for peptide cross-linked dimers and trimers. Addition of benzyl penicillin (300 mug/ml) to growing protoplast cultures did not affect the net amount of PG fragments synthesized but resulted in inhibition of synthesis of dimer and trimer fractions by 27 and 59%, respectively. Failure of penicillin to completely inhibit the accumulation of the dimer fraction was attributed to the presence of atypical forms of dimer. In fact, the supernatant medium of penicillin-treated cultures did not contain detectable amounts of typical peptide cross-linked dimer. The degree of peptide cross-linkage of protoplast PG was at most only 13% of that found in walls isolated from intact streptococci. The relative amounts of monomers, dimers, and trimers synthesized during early and late stages of protoplast growth was approximately the same. Protoplasts synthesized soluble PG fragments in amounts which were of the same order of magnitude as that expected for insoluble PG produced by an equivalent amount of intact streptococci.

Full text

PDF
410

Selected References

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

  1. BRUMFITT W., WARDLAW A. C., PARK J. T. Development of lysozyme-resistance in Micrococcus lysodiekticus and its association with an increased O-acetyl content of the cell wall. Nature. 1958 Jun 28;181(4626):1783–1784. doi: 10.1038/1811783a0. [DOI] [PubMed] [Google Scholar]
  2. Chipman D. M., Pollock J. J., Sharon N. Lysozyme-catalyzed hydrolysis and transglycosylation reactions of bacterial cell wall oligosaccharides. J Biol Chem. 1968 Feb 10;243(3):487–496. [PubMed] [Google Scholar]
  3. Chipman D. M., Sharon N. Mechanism of lysozyme action. Science. 1969 Aug 1;165(3892):454–465. doi: 10.1126/science.165.3892.454. [DOI] [PubMed] [Google Scholar]
  4. Coyette J., Perkins H. R., Polacheck I., Shockman G. D., Ghuysen J. M. Membrane-bound DD-carboxypeptidase and LD-transpeptidase of Streptococcus faecalis ATCC 9790. Eur J Biochem. 1974 May 15;44(2):459–468. doi: 10.1111/j.1432-1033.1974.tb03504.x. [DOI] [PubMed] [Google Scholar]
  5. GHUYSEN J. M., STROMINGER J. L. STRUCTURE OF THE CELL WALL OF STAPHYLOCOCCUS AUREUS, STRAIN COPENHAGEN. II. SEPARATION AND STRUCTURE OF DISACCHARIDES. Biochemistry. 1963 Sep-Oct;2:1119–1125. doi: 10.1021/bi00905a036. [DOI] [PubMed] [Google Scholar]
  6. Ghuysen J. M., Bricas E., Leyh-Bouille M., Lache M., Shockman G. D. The peptide N alpha-(L-alanyl-D-isoglutaminyl)-N epsilon-(D-isoasparaginyl)-L-lysyl-D-alanine and the disaccharide N-acetylglucosaminyl-beta-1,4-N-acetylmuramic acid in cell wall peptidoglycan of Streptococcus faecalis strain ATCC 9790. Biochemistry. 1967 Aug;6(8):2607–2619. doi: 10.1021/bi00860a044. [DOI] [PubMed] [Google Scholar]
  7. Hayashi H., Araki Y., Ito E. Occurrence of glucosamine residues with free amino groups in cell wall peptidoglycan from bacilli as a factor responsible for resistance to lysozyme. J Bacteriol. 1973 Feb;113(2):592–598. doi: 10.1128/jb.113.2.592-598.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Joseph R., Shockman G. D. Autolytic formation of protoplasts (autoplasts) of Streptococcus faecalis 9790: release of cell wall, autolysin, and formation of stable autoplasts. J Bacteriol. 1974 May;118(2):735–746. doi: 10.1128/jb.118.2.735-746.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mirelman D., Bracha R., Sharon N. Role of the penicillin-sensitive transpeptidation reaction in attachment of newly synthesized peptidoglycan to cell walls of Micrococcus luteus. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3355–3359. doi: 10.1073/pnas.69.11.3355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mirelman D., Bracha R., Sharon N. Studies on the elongation of bacterial cell wall peptidoglycan and its inhibition by penicillin. Ann N Y Acad Sci. 1974 May 10;235(0):326–347. doi: 10.1111/j.1749-6632.1974.tb43275.x. [DOI] [PubMed] [Google Scholar]
  11. Rosenthal R. S., Jungkind D., Daneo-Moore L., Shockman G. D. Evidence for the synthesis of soluble peptidoglycan fragments by protoplasts of Streptococcus faecalis. J Bacteriol. 1975 Oct;124(1):398–409. doi: 10.1128/jb.124.1.398-409.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rosenthal R. S., Shockman G. D. Synthesis of peptidoglycan in the form of soluble glycan chains by growing protoplasts (autoplasts) of Streptococcus faecalis. J Bacteriol. 1975 Oct;124(1):419–423. doi: 10.1128/jb.124.1.419-423.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roth G. S., Shockman G. D., Daneo-Moore L. Balanced macromolecular biosynthesis in "protoplasts" of Streptococcus faecalis. J Bacteriol. 1971 Mar;105(3):710–717. doi: 10.1128/jb.105.3.710-717.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SHOCKMAN G. D., KOLB J. J., TOENNIES G. Relations between bacterial cell wall synthesis, growth phase, and autolysis. J Biol Chem. 1958 Feb;230(2):961–977. [PubMed] [Google Scholar]
  15. Shockman G. D., Thompson J. S., Conover M. J. The autolytic enzyme system of Streptococcus faecalis. II. Partial characterization of the autolysin and its substrate. Biochemistry. 1967 Apr;6(4):1054–1065. doi: 10.1021/bi00856a014. [DOI] [PubMed] [Google Scholar]
  16. Taylor A., Das B., van Heijenoort J. Isolement et caractérisation de l'acide N-acétylglucosaminyl-beta-1-4-1, 6-anhydro-N-acétylmuramique. C R Acad Sci Hebd Seances Acad Sci D. 1974 Feb 18;278(8):1127–1129. [PubMed] [Google Scholar]
  17. Ward J. B., Perkins H. R. Peptidoglycan biosynthesis by preparations from Bacillus licheniformis: cross-linking of newly synthesized chains to preformed cell wall. Biochem J. 1974 Jun;139(3):781–784. doi: 10.1042/bj1390781. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES