UNIT 21.16 Monitoring Metalloproteinase Activity Using Synthetic Fluorogenic Substrates
Published Online: 1 NOV 2004
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Troeberg, L. and Nagase, H. 2004. Monitoring Metalloproteinase Activity Using Synthetic Fluorogenic Substrates. Current Protocols in Protein Science. 33:21.16.1–21.16.9.
- Published Online: 1 NOV 2004
- Published Print: AUG 2003
Fluorogenic synthetic substrates are commonly used to monitor the activity of peptidases in vitro. This unit presents a representative protocol that employs (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3-[2,4-dinitrophenyl]-L-2,3-diaminopropionyl)-Ala-Arg-NH2 (Mca-Pro-Leu-Gly~Leu-Dpa-Ala-Arg-NH2) as a substrate to assay matrix metallopeptidases (MMPs). This substrate was first described for the assay of MMP-1, -2 and -3 and it is now widely used as a general MMP substrate. Protocols are given for both stopped-time assays (suitable for assaying MMP activity in a large number of samples) and continuous assays (commonly used when establishing an assay protocol or investigating kinetic aspects of enzyme behavior). Other fluorogenic peptides and protein substrates, together with non-fluorogenic alternatives, are also discussed.