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

Détails

ID Serval
serval:BIB_ED8FF221A81A
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Specific labeling of diaminopimelate: a radioassay for the determination of the peptidoglycan cross-linking index
Périodique
Anal Biochem
Auteur(s)
Roten  C. A., Pagni  M., Margot  P., Touri  F., Karamata  D.
ISSN
0003-2697 (Print)
Statut éditorial
Publié
Date de publication
12/1994
Volume
223
Numéro
2
Pages
208-11
Notes
Comparative Study
Journal Article --- Old month value: Dec
Résumé
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.
Mots-clé
Bacillus subtilis/chemistry Carbon Radioisotopes Cell Wall/chemistry Chromatography, Thin Layer Cross-Linking Reagents Diaminopimelic Acid/*analysis Dinitrofluorobenzene Hydrolysis Peptidoglycan/*chemistry
Pubmed
Web of science
Création de la notice
24/01/2008 14:07
Dernière modification de la notice
20/08/2019 16:15
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