A generic MMP fluorescent substrate, originally prepared as a substrate for fibroblast collagenase. The quenching of the Trp fluorescence in the intact substrate is relieved upon hydrolysis of the P1-P?1 bond, giving rise to a continuously recording fluorescence assay. The hydrolysis rates of the FRET peptide rival or exceed that of its natural protein substrates. This substrate also demonstrates that its water solubility in the typical MMP assay buffer exceeds the Km value for the enzyme, allowing accurate determination of the kinetic parameters.