Click and Release Chemistry for Activity-Based Purification of β-Lactam Targets

Saidbakhrom Saidjalolov (1) , Emmanuelle Braud (1) , Zainab Edoo (2) , Laura Iannazzo (1) , Filippo Rusconi (3) , Margaux Riomet (4) , Antoine Sallustrau (4) , Frédéric Taran (4) , Michel Arthur (2) , Matthieu Fonvielle (2) , Mélanie Etheve-Quelquejeu (1)

1 LCBPT - UMR 8601 - Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques
2 CRC (UMR_S_1138 / U1138) - Centre de Recherche des Cordeliers
3 GQE-Le Moulon - Génétique Quantitative et Evolution - Le Moulon (Génétique Végétale)
4 SCBM - Service de Chimie Bio-Organique et de Marquage

β-Lactams, the cornerstone of antibiotherapy, inhibit multiple and partially redundant targets referred to as transpeptidases or penicillin-binding proteins. These enzymes catalyze the essential cross-linking step of the polymerization of cell wall peptidoglycan. The understanding of the mechanisms of action of β-lactams and of resistance to these drugs requires the development of reliable methods to characterize their targets. Here, we describe an activity-based purification method of β-lactam targets based on click and release chemistry. We synthesized alkyne-carbapenems with suitable properties with respect to the kinetics of acylation of a model target, the Ldtfm L,D-transpeptidase, the stability of the resulting acylenzyme, and the reactivity of the alkyne for the cycloaddition of an azido probe containing a biotin moiety for affinity purification and a bioorthogonal cleavable linker. The probe provided access to the fluorescent target in a single click and release step.

Mots clés: Activity-based probe; Antibiotic; Click and release; Iminosydnone; Peptidoglycan; Activity-based probe

Domaines: Chimie

Saidbakhrom Saidjalolov, Emmanuelle Braud, Zainab Edoo, Laura Iannazzo, Filippo Rusconi, et al.. Click and Release Chemistry for Activity-Based Purification of β-Lactam Targets. Chemistry - A European Journal, 2021, 27 (28), pp.7687-7695. ⟨10.1002/chem.202100653⟩. ⟨hal-03281379⟩