MOCAc-PLGL(Dpa)AR |
| Catalog No.GC17832 |
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 140430-53-1
Sample solution is provided at 25 µL, 10mM.
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We have retained the Pro-Leu-Gly-Lcu sequence, but labelled the N-terminus with the highly fluorescent (7-methoxycoumarin-4-yl) acetyl (Mea) group. The quenching Dnp group has been inserted with the novel N3-Dnp-t_-2.3-diaminopropionic acid (Dpa) in the P2' position1, as the matrix metalloproteinases favour aromatic side chains at P2'2, 3.
In the unquenched peptide Mca-Pro-Leu, the fluorescence has an excitation maximum at 328 nm and the emission is centered around 393 nm. When the fluorescence emissions were compared at a concentration of 1 ,µM, Mca-Pro-Leu was 130 times more fluorescent than Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2.
3.3 CIeavuge of Mea-Pro-Leu-Gus)-Lrlt-Dpu-AMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2, is the most sensitive substrate so far described for continuous assays of the matrix metalloproteinases. Not only are the values of Kcut/Km, higher than those reported previously. but the cleavage product is more readily detectable, as the Mea group is both more fluorescent than tryptophan and more efficiently quenched by the Dnp group. With the exception of collegenase, continuous assays can be made with the new substrate at enzyme concentrations similar to those required in assays with their preferred macromolecular substrates. Mca-Pro- Leu-Gly-Leu-Dpa-Ala-Arg-NH2 is suitable therefore for the assessment of total matrix metalloproteinase activity in crude preparations. Further work to develop peptides that are specifically cleaved by the individual enzymes will permit their measurement under these conditions3.
Reference:
1. Schectcr, I, and Beryer. A. (lYG7) Biochem. Biophyt;. Rcs, Commun. 27, 157-161
2. Nctzcl-Arnctt, s.. Mallya. SK., Nugasc. H.. Birkedal-Hansen, H. and Van Wurt. H.E. (1991) Anal. Biochcm. 195. 86-92.
3. Netzel-Arnctt, S., Fields. G., Birkedal-Hunscn. H. and Vai Wart. H.E. (1991) J. Biol. Chem. 266. 6747-6755
4. C.G.Knight, F. Willenbrock and G. Murphy. A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases. FEDS. Volume 296, number 3, 263-266
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