A versatile assay for gelatinases using succinylated gelatin

A spectrophotometric assay using succinylated gelatin as substrate is descr ibed for measuring the catalytic activity of gelatinases. The assay is base d on measurement of primary amines exposed as a result of hydrolysis of the substrate by gelatinases. Comparison of hydrolysis by matrix metalloprotei nase (MMP) 1, 2, 3, 7, 9 indicated that succinylated gelatin was primarily digested by MMP-2 and -9. The assay is rapid (< 60 min), specific, suitable for measuring gelatinolytic activity of enzymes and high volume screening of MMP-2 and -9 inhibitors. Sensitivity of the assay is comparable to that of gelatin zymography, under similar experimental conditions. Thus, the ass ay combines ease and rapidity of assays based on synthetic peptide substrat es with specificity of the gelatin zymography technique. (C) 2000 Elsevier Science B.V./International Society of Matrix Biology. All rights reserved.