Membrane Type-1 Matrix Metalloprotease and Stromelysin-3 Cleave More Efficiently Synthetic Substrates Containing Unusual Amino Acids in Their P1′ Positions
| Autor: | Vincent Dive, Rama Kannan, Athanasios Yiotakis, Artur Mucha, Philippe Cuniasse, Paul Basset, Fabrice Beau |
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| Rok vydání: | 1998 |
| Předmět: |
Dansyl Compounds
chemistry.chemical_classification Binding Sites Matrix Metalloproteinases Membrane-Associated Stereochemistry Metalloendopeptidases Substrate (chemistry) Phenylalanine Peptide Cell Biology Biochemistry Substrate Specificity Amino acid Zinc Residue (chemistry) Chromogenic Compounds chemistry Matrix Metalloproteinase 11 Cleave Metalloproteins Amino Acids Leucine Oligopeptides Molecular Biology Cysteine |
| Zdroj: | Journal of Biological Chemistry. 273:2763-2768 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.273.5.2763 |
| Popis: | The influence of the substrate P1′ position on the specificity of two zinc matrix metalloproteases, membrane type-1 matrix metalloprotease (MT1-MMP) and stromelysin-3 (ST3), was evaluated by synthesizing a series of fluorogenic substrates of general formula dansyl-Pro-Leu-Ala-Xaa-Trp-Ala-Arg-NH2, where Xaa in the P1′ position represents unusual amino acids containing either long arylalkyl or alkyl side chains. Our data demonstrate that both MT1-MMP and ST3 cleave substrates containing in their P1′ position unusual amino acids with extremely long side chains more efficiently than the corresponding substrates with natural phenylalanine or leucine amino acids. In this series of substrates, the replacement of leucine by S-para-methoxybenzyl cysteine increased the k cat/K m ratio by a factor of 37 for MT1-MMP and 9 for ST3. The substrate with aS-para-methoxybenzyl cysteine residue in the P1′ position displayed ak cat/K m value of 1.59 106 m −1 s−1 and 1.67 104 m −1 s−1, when assayed with MT1-MMP and ST3, respectively. This substrate is thus one of the most rapidly hydrolyzed substrates so far reported for matrixins, and is the first synthetic peptide efficiently cleaved by ST3. These unexpected results for these two matrixins suggest that extracellular proteins may be cleaved by matrixins at sites containing amino acids with unusual long side chains, like those generatedin vivo by some post-translational modifications. |
| Databáze: | OpenAIRE |
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