An Activity-Based Probe for Studying Crosslinking in Live Bacteria.
Angew Chem Int Ed Engl
; 54(36): 10492-6, 2015 Sep 01.
Article
em En
| MEDLINE
| ID: mdl-26204841
ABSTRACT
Penicillin-binding proteins (PBPs) catalyze the crosslinking of peptidoglycan (PG), an essential process for bacterial growth and survival, and a common antibiotic target. Yet, despite its importance, little is known about the spatiotemporal aspects of crosslinkinglargely because of a lack of experimental tools for studying the reaction in live bacteria. Here we introduce such a tool an activity-based probe that enables visualization and relative quantitation of crosslinking in vivo. In Staphylococcus aureus, we show that fluorescent mimics of the natural substrate of PBPs (PG stem peptide) are covalently incorporated into the cell wall, installing fluorophores in place of natural crosslinks. These fluorescent stem peptide mimics (FSPMs) are selectively recognized by a single PBP in S. aureus PBP4. Thus, we were able to use FSPM pulse-labeling to localize PBP4 activity in live cells, showing that it is recruited to the septum in a manner dependent on wall teichoic acid.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Staphylococcus aureus
/
Sondas Moleculares
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article