Matrix metalloproteinases (MMPs...

A selective, sensitive, and convenient assay for human collagenase is required because of its im-plication in diseases such as rheumatoid arthritis, osteoarthritis, and tumors. Here, a novel assay for human collagenase activity is described in which enzymatic degradation of native collagen or acetyl peptide is determined by using a fluorogenic reaction for oligopeptides. The oligopeptides are quantified spectrofluorometrically with either 3,4-dihydroxyphenylacetic acid or 1,2-dihy-droxybenzen reaction in the presence of sodium periodate and sodium borate (pH 7- 8). These reactions can selectively convert N-terminal Gly-containing oligopeptides and N-terminal Ile-con-taining oligopeptides to fluorescence (FL) compounds, respectively, but not proteins, acetyl pep-tides or amino acids. Under optimized conditions using 1.65 μM collagen IV or 1.5 mM Ac-GPQGI-AGQ as substrates, this assay exhibits a proportional relationship between FL intensities and hu-man collagenase-3 (MMP-13) concentrations. It can assay endogenous collagenase activities i