Περίληψη:
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 P1′ position, and a tritium radioactive label for the sensitive detection of MMP covalent adducts by radioimaging 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. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Συγγραφείς:
Nury, C.
Czarny, B.
Cassar-Lajeunesse, E.
Georgiadis, D.
Bregant, S.
Dive, V.
Λέξεις-κλειδιά:
collagenase 3; gelatinase A; gelatinase B; macrophage elastase; matrix metalloproteinase; matrix metalloproteinase 14; neutrophil collagenase; stromelysin; tritium, article; chemical modification; controlled study; cross linking; enzyme mechanism; photoaffinity labeling; priority journal; radioactivity; radiography, Azides; Azirines; Catalytic Domain; Cross-Linking Reagents; Humans; Light; Matrix Metalloproteinases; Molecular Probe Techniques; Photoaffinity Labels
