Purification of Active Matrix Metalloproteinase Catalytic Domains and Its Use for Screening of Specific Stromelysin-3 Inhibitors
| Autor: | Vincent Dive, John G. Kochins, Agnès Noël, Rama Kannan, Susan P. Manly, Paul Basset, Mostafa El Fahime, Marc Ruff, Marie-Christine Rio, Jean-Michel Foidart, Isabelle Stoll |
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| Rok vydání: | 1999 |
| Předmět: |
Matrix Metalloproteinases
Membrane-Associated Matrix metalloproteinase inhibitor Drug Evaluation Preclinical Antineoplastic Agents CHO Cells In Vitro Techniques Matrix metalloproteinase Inclusion bodies Mice Matrix Metalloproteinase 10 Matrix Metalloproteinase 11 Catalytic Domain Cricetinae Escherichia coli Animals Humans Protease Inhibitors Matrilysin DNA Primers chemistry.chemical_classification Metalloproteinase Base Sequence Metalloendopeptidases Substrate (chemistry) Molecular biology Recombinant Proteins In vitro Kinetics Enzyme chemistry Biochemistry Matrix Metalloproteinase 7 Biotechnology |
| Zdroj: | Protein Expression and Purification. 16:76-83 |
| ISSN: | 1046-5928 |
| DOI: | 10.1006/prep.1999.1068 |
| Popis: | The matrix metalloproteinase (MMP) stromelysin-3 (ST3) has been shown to be involved in malignant tumor progression and therefore represents an attractive therapeutical target. In order to screen for ST3 synthetic inhibitors, we have produced and purified the catalytic domain of ST3, matrilysin, stromelysin-2, and membrane type-1 MMP from inclusion bodies in a bacterial system. Our strategy allowed the purification of MMPs directly in the active form, thereby avoiding in vitro activation. A total of 140,000 synthetic compounds from the Bristol-Myers Pharmaceutical Research Institute chemical deck were tested, using a substrate-based colorimetric enzymatic assay, in which ST3 activity was evaluated through its ability to cleave and inactivate alpha-1 proteinase inhibitor. One ST3 inhibitor belonging to the cephalosporin family of antibiotics was thereby identified. |
| Databáze: | OpenAIRE |
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