Anal Biochem. 1994 Dec;223(2):208-11.

Specific labeling of diaminopimelate: a radioassay for the determination of the peptidoglycan cross-linking index.

Roten CA, Pagni M, Margot P, Touri F, Karamata D.

Source

Institut de génétique et de biologie microbiennes, Lausanne, Switzerland.

Abstract

A radioassay for the determination of the peptidoglycan cross-linking index (CLI) was devised. It is based on specific radioactive labeling of diaminopimelic acid (DAP) by diverting [14C]aspartate into the DAP pathway, while inhibiting incorporation of label into other cell wall components. Purified [14C]DAP-labeled cell walls were treated with fluorodinitrobenzene, hydrolyzed, and chromatographed by TLC. The radioactivity in well-separated mono dinitrophenyl-diaminopimelate (DNP-DAP) and DAP spots was counted and the CLI was determined from the ratio of DAP to the total of mono DNP-DAP and DAP counts. The method, suitable for bacteria such as Bacillus subtilis unable to incorporate exogenous DAP, can be applied to other systems. A CLI of 50.8 +/- 1.3% and 55.5 +/- 0.9% was obtained for B. subtilis 168 cells growing exponentially in rich and minimal medium, respectively. Comparison of these to results previously obtained on B. subtilis suggested the existence of a hitherto unreported peptidoglycan endopeptidase activity.

PMID:: 7887465; [PubMed - indexed for MEDLINE]

Publication Types, MeSH Terms, Substances

Publication Types

Comparative Study

MeSH Terms

Substances

LinkOut - more resources

Full Text Sources

Molecular Biology Databases

1-FLUORO-2,4-DINITROBENZENE - HSDB

Supplemental Content

Save items

Related citations in PubMed

Characterization of cross-linking of cell walls of Bacillus subtilis by a combination of magic-angle spinning NMR and gas chromatography-mass spectrometry of both intact and hydrolyzed 13C- and 15N-labeled cell-wall peptidoglycan. [J Biol Chem. 1991]
Characterization of cross-linking of cell walls of Bacillus subtilis by a combination of magic-angle spinning NMR and gas chromatography-mass spectrometry of both intact and hydrolyzed 13C- and 15N-labeled cell-wall peptidoglycan.
Forrest TM, Wilson GE, Pan Y, Schaefer J. J Biol Chem. 1991 Dec 25; 266(36):24485-91.
Analysis of peptidoglycan structure from vegetative cells of Bacillus subtilis 168 and role of PBP 5 in peptidoglycan maturation. [J Bacteriol. 1999]
Analysis of peptidoglycan structure from vegetative cells of Bacillus subtilis 168 and role of PBP 5 in peptidoglycan maturation.
Atrih A, Bacher G, Allmaier G, Williamson MP, Foster SJ. J Bacteriol. 1999 Jul; 181(13):3956-66.
N-acetylmuramoyl-L-alanine amidase assay based on specific radioactive labeling of muropeptide L-alanine: quantitation of the enzyme activity in the autolysin deficient Bacillus subtilis 168, flaD strain. [Anal Biochem. 1991]
N-acetylmuramoyl-L-alanine amidase assay based on specific radioactive labeling of muropeptide L-alanine: quantitation of the enzyme activity in the autolysin deficient Bacillus subtilis 168, flaD strain.
Margot P, Roten CA, Karamata D. Anal Biochem. 1991 Oct; 198(1):15-8.
Review Another brick in the wall. [Trends Microbiol. 2007]
Review Another brick in the wall.
Pavelka MS Jr. Trends Microbiol. 2007 Apr; 15(4):147-9. Epub 2007 Feb 28.
Review The role of peptidoglycan structure and structural dynamics during endospore dormancy and germination. [Antonie Van Leeuwenhoek. 1999]
Review The role of peptidoglycan structure and structural dynamics during endospore dormancy and germination.
Atrih A, Foster SJ. Antonie Van Leeuwenhoek. 1999 May; 75(4):299-307.

See reviews... See all...

Cited by 1 PubMed Central article

Analysis of the peptidoglycan structure of Bacillus subtilis endospores. [J Bacteriol. 1996]
Analysis of the peptidoglycan structure of Bacillus subtilis endospores.
Popham DL, Helin J, Costello CE, Setlow P. J Bacteriol. 1996 Nov; 178(22):6451-8.

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Compound (MeSH Keyword)
PubChem chemical compound records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

Specific labeling of diaminopimelate: a radioassay for the determination of the ...
Specific labeling of diaminopimelate: a radioassay for the determination of the peptidoglycan cross-linking index.
Anal Biochem. 1994 Dec ;223(2):208-11.

PubMed
Prospective assessment of risks for cervicomedullary-junction compression in inf...
Prospective assessment of risks for cervicomedullary-junction compression in infants with achondroplasia.
Am J Hum Genet. 1995 Mar ;56(3):732-44.

PubMed
[Neuroses--myths, models, facts. A contribution to the genesis of neurotic distu...
[Neuroses--myths, models, facts. A contribution to the genesis of neurotic disturbances].
Bibl Psychiatr. 1976 ;(153):1-111.

PubMed
Crohn's disease with Behçet's syndrome like appearance: a case report.
Crohn's disease with Behçet's syndrome like appearance: a case report.
Z Gastroenterol. 1994 Nov ;32(11):642-4.

PubMed
Hemophilia with factor VIII and factor IX inhibitors, incidence, bleeding proble...
Hemophilia with factor VIII and factor IX inhibitors, incidence, bleeding problems and management.
Southeast Asian J Trop Med Public Health. 1993 ;24 Suppl 1:106-12.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: