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Yet, despite its importance, little is known about the spatiotemporal aspects of crosslinking\u2014largely because of a lack of experimental tools for studying the reaction in live bacteria. Here we introduce such a tool: an activity\u2010based 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\u2010labeling to localize PBP4 activity in live cells, showing that it is recruited to the septum in a manner dependent on wall teichoic acid.<\/jats:p>","DOI":"10.1002\/anie.201503869","type":"journal-article","created":{"date-parts":[[2015,7,24]],"date-time":"2015-07-24T01:07:32Z","timestamp":1437700052000},"page":"10492-10496","source":"Crossref","is-referenced-by-count":36,"title":["An Activity\u2010Based Probe for Studying Crosslinking in Live 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