A Pan Photoaffinity Probe for Detecting Active Forms of Matrix Metalloproteinases

作者：Nury Catherine; Czarny Bertrand; Cassar Lajeunesse Evelyne; Georgiadis Dimitris; Bregant Sarah; Dive Vincent^*

来源：ChemBioChem, 2013, 14(1): 107-114.

DOI：10.1002/cbic.201200583

• activity-based probes
• diazirines
• matrix metalloproteases
• metalloenzymes
• photoaffinity labeling

A photoaffinity probe based on the scaffold of a potent broad-spectrum phosphinic peptide inhibitor of matrix metalloproteinases (MMPs) has been developed. A photolabile diazirine group for covalent modification of MMP active forms was incorporated at the P-1%26apos; position, and a tritium radioactive label for the sensitive detection of MMP covalent adducts by radio-imaging was attached. The probe was characterized on seven catalytic domains of human MMPs (MMP-2, -3, -8, -9, -12, -13 and -14) and was found to display nanomolar affinities towards this set of MMPs, covalently modifying them with crosslinking yields varying from 12 to 58%, thus leading to highly sensitive detection of these MMPs. In a complex proteome complemented with four recombinant MMPs (MMP-2, -9, -12 and -13), this probe enabled their simultaneous detection with a threshold of few femtomoles and low background labelling. Those properties should make this new pan-activity-based MMP probe a valuable tool for the detection of MMP active forms from biological fluids or tissue extracts.

• 出版日期2013-1-2
• 单位中国地震局