Ca2+ and Zn2+ binding properties of peptide substrates of vertebrate collagenase, MMP-1
| Autor: | Vettai S. Ananthanarayanan, Nataraj N Vettakkorumakankav |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Circular dichroism
Stereochemistry Biophysics Peptide Matrix Metalloproteinase Inhibitors Biochemistry Substrate Specificity Structural Biology Escherichia coli medicine Humans Amino Acid Sequence Collagenases Binding site Molecular Biology Peptide sequence chemistry.chemical_classification Oligopeptide Binding Sites Tissue Inhibitor of Metalloproteinase-1 biology Circular Dichroism Hydrolysis Active site Recombinant Proteins Zinc Enzyme chemistry biology.protein Collagenase Calcium Matrix Metalloproteinase 1 Oligopeptides Plasmids medicine.drug |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology. 1432:356-370 |
| ISSN: | 0167-4838 |
| DOI: | 10.1016/s0167-4838(99)00111-9 |
| Popis: | To understand the role of Ca(2+) in vertebrate in the structure and action of collagenase, we have examined peptides that interact with recombinant human fibroblast collagenase for their affinities towards Ca(2+) and Zn(2+) in a non-polar solvent. Two of the peptides, GPQGIAGQ and GNVGLAGA, had sequences in collagen which are, respectively, cleaved and not cleaved by collagenase. A third peptide, PSYFLNAG, had a collagenase-cleaved sequence in ovostatin, a globular protein substrate. Peptides TVGCEECTV and CLPREPGL were derived from TIMP-1; the former competitively inhibits collagenase while the latter does not. The relative rates of hydrolysis of the peptides by collagenase had the order GPQGIAGQ>PSYFLNAG>GNVGLAGA. Circular dichroism spectral data in trifluoroethanol showed that while the TIMP control peptide, CLPREPGL, bound only Zn(2+), the other four peptides bound both Ca(2+) and Zn(2+) with definite stoichiometries. Ca(2+) could displace Zn(2+) in the substrate peptides while Zn(2+) displaced Ca(2+) in the TIMP peptide. GPQGIAGQ, PSYFLNAG and TVGCEECTV formed peptide:Ca(2+):Zn(2+) ternary complexes. Our results suggest that both collagen and globular protein substrates of collagenase may bind Ca(2+) and Zn(2+) in the enzyme's active site. This, in turn, may account for the known importance of the non-catalytic Ca(2+) and Zn(2+) in collagenase activity. |
| Databáze: | OpenAIRE |
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