Expression of Human Pro-Matrix Metalloproteinase 3 that Lacks the N-terminal 34 Residues in Escherichia coli: Autoactivation and Interaction with Tissue Inhibitor of Metalloproteinase 1 (TIMP-1)
| Autor: | Chen-Chen Kan, Ko Suzuki, Keith Brew, Hideaki Nagase, Wen Hung, Michael R. Gehring |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Matrix Metalloproteinase 3
Recombinant Fusion Proteins Clinical Biochemistry Gene Expression Matrix metalloproteinase medicine.disease_cause Biochemistry chemistry.chemical_compound Escherichia coli medicine Humans Protease Inhibitors Protein precursor Molecular Biology Sequence Deletion chemistry.chemical_classification Enzyme Precursors Metalloproteinase Tissue Inhibitor of Metalloproteinase-1 Tissue inhibitor of metalloproteinase Cell biology Amino acid Enzyme Activation EGTA chemistry |
| Zdroj: | bchm. 379:185-192 |
| ISSN: | 1437-4315 1431-6730 |
| DOI: | 10.1515/bchm.1998.379.2.185 |
| Popis: | Human pro-matrix metalloproteinase 3 (proMMP-3) lacking the N-terminal 34 amino acids and the C-terminal hemopexin-like domain was expressed in E. coli and used to investigate the process of proenzyme activation and its interaction with an endogenous inhibitor TIMP-1 during activation. The truncated precursor was purified from the E. coli extract in the presence of 5mM EGTA. The active 23.5 kDa form was generated simply by exposure to Ca2+ and Zn2+ but not either by Ca2+ alone or by Zn2+ alone. The rate of MMP-3(deltaC) formation was concentration dependent, indicating that autoactivation is a bimolecular reaction. The truncated precursor was able to interact with the N-terminal domain of TIMP-1 without losing the 48 residue-long propeptide. However, upon a longer incubation, the propeptide was slowly processed, indicating that the association of the N-terminally truncated proMMP-3 with TIMP-1 is weaker than that of the fully activated MMP-3 and TIMP-1. These results indicate that the expression of MMP activities is regulated by endogenous inhibitor TIMPs during their activation processes which provide an additional control mechanism of extracellular matrix breakdown. |
| Databáze: | OpenAIRE |
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