Analysis of Gram-negative Bacteria Peptidoglycan by Ultra-performance Liquid Chromatography.
Cell Wall
Crosslink
Muramidase
Muropeptide
Peptidoglycan
UPLC
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Oct 2020
05 Oct 2020
Historique:
received:
12
06
2020
revised:
20
08
2020
accepted:
21
09
2020
entrez:
4
3
2021
pubmed:
5
3
2021
medline:
5
3
2021
Statut:
epublish
Résumé
Bacteria are surrounded by a protective peptidoglycan cell wall. Provided that this structure and the enzymes involved are the preferred target for our most successful antibiotics, determining its structural and chemical complexity is of the highest interest. Traditionally, high-performance liquid chromatography (HPLC) analyses have been performed, but these methods are very time consuming in terms of sample preparation and chromatographic separation. Here we describe an optimized method for preparation of Gram-negative bacteria peptidoglycan and its subsequent analysis by ultra-performance liquid chromatography (UPLC). The use of UPLC in peptidoglycan analyses provides a dramatic reduction of the sample volume and hands-on time required and, furthermore, permits in-line mass spectrometry (MS) of the UPLC resolved muropeptides, thus facilitating their identification. This method improves our capability to perform high throughput analysis to better understand the cell-wall biology.
Identifiants
pubmed: 33659436
doi: 10.21769/BioProtoc.3780
pii: e3780
pmc: PMC7842696
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e3780Informations de copyright
Copyright © The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no conflict of interest or competing interest.
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