A Pan Photoaffinity Probe for Detecting Active Forms of Matrix Metalloproteinases
| Autor: | Bertrand Czarny, Sarah Bregant, Dimitris Georgiadis, Vincent Dive, Catherine Nury, Evelyne Cassar-Lajeunesse |
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| Rok vydání: | 2012 |
| Předmět: |
Radioactive Label
Azides Light Molecular Probe Techniques Photoaffinity Labels Matrix metalloproteinase Biochemistry Adduct law.invention chemistry.chemical_compound law Catalytic Domain Humans Molecular Biology Photoaffinity labeling Azirines Organic Chemistry Matrix Metalloproteinases Cross-Linking Reagents chemistry Covalent bond Diazirine Proteome Recombinant DNA Molecular Medicine |
| Zdroj: | ChemBioChem. 14:107-114 |
| ISSN: | 1439-4227 |
| DOI: | 10.1002/cbic.201200583 |
| Popis: | 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) ' 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. |
| Databáze: | OpenAIRE |
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