Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Feb;157(2):398–404. doi: 10.1128/jb.157.2.398-404.1984

Biosynthesis of peptidoglycan in Gaffkya homari: processing of nascent glycan by reactivated membranes.

C Bardin, R K Sinha, E Kalomiris, F C Neuhaus
PMCID: PMC215261  PMID: 6693347

Abstract

Membranes from Gaffkya homari reactivated by freezing and thawing were used to study the processing events involved in the assembly of both sodium dodecyl sulfate (SDS)-insoluble peptidoglycan (PG) and SDS-soluble PG. The ability to reactivate membranes for the synthesis of these polymers provided an opportunity to monitor those events that are not influenced by wall-linked PG. In G. homari, processing for the formation of cross-links requires the selective actions of DD-carboxypeptidase, LD-carboxypeptidase, and NE-(DAla)-Lys transpeptidase. Time courses of cross-link formation, as measured by the amounts of amidated bisdisaccharide peptide dimer and nonamidated bisdisaccharide peptide dimer, showed a lack of correlation with those for the synthesis of SDS-insoluble PG. SDS-soluble PG, which is significantly cross-linked when synthesized in the absence of penicillin G, was a precursor of the SDS-insoluble PG. In the presence of penicillin G, un-cross-linked SDS-soluble PG was synthesized. This PG was also utilized and processed for the synthesis of cross-linked SDS-insoluble PG after removal of the beta-lactam. This protocol provided a method for separating stages in the synthesis and elongation of PG from those involved in processing. Cross-linkage in the various PG fractions ranged from 0 to 19% in SDS-soluble PG and from 2 to 24% in SDS-insoluble PG. Thus, the results indicated that there is no direct correlation between SDS insolubility and the degree of cross-linkage. Instead, they suggested that additional features may contribute to the insolubility of PG in SDS.

Full text

PDF
398

Selected References

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

  1. Blumberg P. M., Yocum R. R., Willoughby E., Strominger J. L. Binding of (14C)penicillin G to the membrane-bound and the purified D-alanine carboxypeptidases from Bacillus stearothermophilus and Bacillus subtilis and its release. J Biol Chem. 1974 Nov 10;249(21):6828–6835. [PubMed] [Google Scholar]
  2. Carpenter C. V., Goyer S., Neuhaus F. C. Steric effects on penicillin-sensitive peptidoglycan synthesis in a membrane-wall system Gaffkya homari. Biochemistry. 1976 Jul 13;15(14):3146–3152. doi: 10.1021/bi00659a031. [DOI] [PubMed] [Google Scholar]
  3. Gmeiner J. Identification of peptide-cross-linked trisdisaccharide peptide trimers in murein of Escherichia coli. J Bacteriol. 1980 Jul;143(1):510–512. doi: 10.1128/jb.143.1.510-512.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hammes W. P. Biosynthesis of peptidoglycan in Gaffkya homari. The mode of action of penicillin G and mecillinam. Eur J Biochem. 1976 Nov 1;70(1):107–113. doi: 10.1111/j.1432-1033.1976.tb10961.x. [DOI] [PubMed] [Google Scholar]
  5. Hammes W. P., Kandler O. Biosynthesis of peptidoglycan in Gaffkya homari. The incorporation of peptidoglycan into the cell wall and the direction of transpeptidation. Eur J Biochem. 1976 Nov 1;70(1):97–106. doi: 10.1111/j.1432-1033.1976.tb10960.x. [DOI] [PubMed] [Google Scholar]
  6. Hammes W. P., Neuhaus F. C. Biosynthesis of peptidoglycan in Gaffkya homari: role of the peptide subunit of uridine diphosphate-N-acetylmuramyl-pentapeptide. J Bacteriol. 1974 Oct;120(1):210–218. doi: 10.1128/jb.120.1.210-218.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hammes W. P., Seidel H. The activities in vitro of DD-carboxypeptidase and LD-carboxypeptidase of Gaffkya homari during biosynthesis of peptidoglycan. Eur J Biochem. 1978 Mar;84(1):141–147. doi: 10.1111/j.1432-1033.1978.tb12150.x. [DOI] [PubMed] [Google Scholar]
  8. Hammes W. P. The LD-carboxypeptidase activity in Gaffkya homari. The target of the action of D-amino acids or glycine on the formation of wall-bound peptidoglycan. Eur J Biochem. 1978 Nov 15;91(2):501–507. doi: 10.1111/j.1432-1033.1978.tb12703.x. [DOI] [PubMed] [Google Scholar]
  9. Kalomiris E., Bardin C., Neuhaus F. C. Biosynthesis of peptidoglycan in Gaffkya homari: reactivation of membranes by freeze-thawing in the presence and absence of walls. J Bacteriol. 1982 May;150(2):535–544. doi: 10.1128/jb.150.2.535-544.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mett H., Bracha R., Mirelman D. Soluble nascent peptidoglycan in growing Escherichia coli cells. J Biol Chem. 1980 Oct 25;255(20):9884–9890. [PubMed] [Google Scholar]
  11. Mirelman D., Nuchamowitz Y. Biosynthesis of peptidoglycan in Pseudomonas aeruginosa. 1. The incorporation of peptidoglycan into the cell wall. Eur J Biochem. 1979 Mar;94(2):541–548. doi: 10.1111/j.1432-1033.1979.tb12923.x. [DOI] [PubMed] [Google Scholar]
  12. Nakel M., Ghuysen J. M., Kandler O. Wall peptidoglycan in Aerococcus viridans strains 201 Evans and ATCC 11563 and in Gaffkya homari strain ATCC 10400. Biochemistry. 1971 May 25;10(11):2170–2175. doi: 10.1021/bi00787a033. [DOI] [PubMed] [Google Scholar]
  13. Neuhaus F. C., Tobin C. E., Ahlgren J. A. Membrane-wall interrelationship in Gaffkya homari: sulfhydryl sensitivity and heat lability of nascent peptidoglycan incorporation into walls. J Bacteriol. 1980 Jul;143(1):112–119. doi: 10.1128/jb.143.1.112-119.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reisinger P., Seidel H., Tschesche H., Hammes W. P. The effect of nisin on murein synthesis. Arch Microbiol. 1980 Oct;127(3):187–193. doi: 10.1007/BF00427192. [DOI] [PubMed] [Google Scholar]
  15. Reynolds P. E., Shepherd S. T., Chase H. A. Identification of the binding protein which may be the target of penicillin action in Bacillus megaterium. Nature. 1978 Feb 9;271(5645):568–570. doi: 10.1038/271568a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Thorpe S. J., Perkins H. R. Deoxycholate enhancement of an intermediate of peptidoglycan synthesis in Micrococcus luteus. FEBS Lett. 1979 Sep 1;105(1):151–154. doi: 10.1016/0014-5793(79)80906-0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES