Simultaneous analyses of clostridium collagenase and neutral proteinase activities with a single synthetic substrate.

K Nomura, Kohji Nomura

Index: Anal. Biochem. 107(1) , 96-102, (1980)

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Abstract

Two simple methods are presented for simultaneously detecting the activities of both Clostridium histolyticum collagenase and a neutral proteinase that contaminates commercial preparations of collagenase, using a single common substrate carbobenzoxy-Gly-Pro-Leu-Gly-Pro. This pentapeptide is cleaved only once either at the Leu-Gly bond by collagenase or at the Pro-Leu bond by the neutral proteinase. Both products, Gly-Pro and Leu-Gly-Pro, can be analyzed not only semiquantitatively by thin-layer chromatography on cellulose sheets, but also quantitatively by direct application of the digest to an automatic amino aicd analyzer for the calculation of initial velocities. These methods have an advantage over the conventional caseinolysis assay in the time required for incubation. A tetrapeptide, carbobenzoxy-Gly-Pro-Leu-Gly, is also cleaved by the neutral proteinase, but less effectively than the pentapeptide. It is proposed that this proteinase has thermolysin-like specificity.