Proximity‐Induced Transfer of a Mass Tag Enables Direct Profiling of Active Matrix Metalloproteases
Abstract
ABSTRACT Conventional activity‐based probes in activity‐based protein profiling (ABPP) require enrichment or reporter tags for detection, which limits sensitivity and multiplexing. Here, we present an enrichment‐free chemoproteomic approach that enables direct mass spectrometric detection by Matrix‐Assisted Laser Desorption/Ionization (MALDI) of active proteases. An active‐site–directed affinity probe transfers, through a proximity‐induced reaction, a MALDI‐detectable α‐cyano‐4‐hydroxycinnamic acid (CHCA) tag exclusively to catalytically active forms of matrix metalloproteases (MMPs). The CHCA label enhances ionization efficiency and markedly improves signal‐to‐noise ratios, allowing confident identification of CHCA‐labelled peptides under discriminating analytical conditions. Each active metalloprotease is thereby, associated with a distinct set of CHCA signature peptides, defining its activity fingerprint. This workflow achieves multiplexed and quantitative activity profiling of MMPs, directly in complex proteomes. This design expands ABPP into the mass spectrometry domain and establishes a robust platform for activity‐based enzyme detection.
Article Details
Authors (21)
Lomane Berthy
CEA, INRAE, Médicaments et Technologies pour La Santé (MTS), SIMoS Université Paris‐Saclay Gif‐sur‐Yvette France
Ugo Pasco
IBMM, Univ. Montpellier, ENSCM CNRS Montpellier France
Mylene Sejalon‐Cipolla
IBMM, Univ. Montpellier, ENSCM CNRS Montpellier France
Pierrick Bruyat
CEA, INRAE, Médicaments et Technologies pour La Santé (MTS), SIMoS Université Paris‐Saclay Gif‐sur‐Yvette France
Monika Kaminska
Carole Malgorn
CEA, INRAE, Médicaments et Technologies pour La Santé (MTS), SIMoS Université Paris‐Saclay Gif‐sur‐Yvette France
Fabrice Beau
CEA, INRAE, Médicaments et Technologies pour La Santé (MTS), SIMoS Université Paris‐Saclay Gif‐sur‐Yvette France
Manca Osolin
National Institute of Biology Department of Genetic Toxicology and Cancer Biology, 121 Večna pot Ljubljana Slovenia
Metka Novak
National Institute of Biology Department of Genetic Toxicology and Cancer Biology, 121 Večna pot Ljubljana Slovenia
Barbara Breznik
National Institute of Biology Department of Genetic Toxicology and Cancer Biology, 121 Večna pot Ljubljana Slovenia
Dimitris Georgiadis
Department of Chemistry, Laboratory of Organic Chemistry University of Athens, Panepistimiopolis, Zografou Athens Greece
Isabelle Correia
Sorbonne Université, Ecole Normale Supérieure, PSL University, CNRS Chimie Physique et Chimie du Vivant CPCV Paris France
Emeline Groult
IRMB, University of Montpellier INSERM, CHU Montpellier Montpellier France
Olivier Lequin
Sarah Bregant
CEA, INRAE, Médicaments et Technologies pour La Santé (MTS), SIMoS Université Paris‐Saclay Gif‐sur‐Yvette France
Yves‐Marie Pers
University of Ljubljana Biotechnical Faculty Ljubljana Slovenia
Daniele Noel
University of Ljubljana Biotechnical Faculty Ljubljana Slovenia
Robert Thai
Université Paris-Saclay, CEA, INRAE, Médicaments et Technologies pour la Santé (MTS), SIMoS
Gilles Subra
IBMM, Univ. Montpellier, ENSCM CNRS Montpellier France
Sonia Cantel
IBMM, Univ. Montpellier, ENSCM CNRS Montpellier France
Laurent Devel