Click and Release Chemistry for Activity-Based Purification of β-Lactam Targets |
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Authors: | Saidbakhrom Saidjalolov Dr Emmanuelle Braud Dr Zainab Edoo Dr Laura Iannazzo Dr Filippo Rusconi Dr Margaux Riomet Dr Antoine Sallustrau Dr Frédéric Taran Dr Michel Arthur Dr Matthieu Fonvielle Prof Dr Mélanie Etheve-Quelquejeu |
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Institution: | 1. Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601 CNRS, Université de Paris, 45, rue des saints-pères, Paris, 75006 France;2. INSERM UMRS 1138, Sorbonne Universités, UPMC Univ Paris 06, Sorbonne Paris Cité, Université de Paris, Centre de recherche des Cordeliers, Paris, 75006 France;3. PAPPSO, Université Paris-Saclay, INRAE, CNRS, AgroParisTech GQE - Le Moulon, Gif-sur-Yvette, 91190 France;4. Université Paris Saclay, CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS), SCBM, Gif-sur-Yvette, 91191 France |
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Abstract: | 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. |
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Keywords: | Activity-based probe Antibiotic Click and release Iminosydnone Peptidoglycan |
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