PURIFICATION OF TADPOLE COLLAGENASE AND CHARACTERIZATION USING COLLAGEN AND SYNTHETIC SUBSTRATES

Tadpole collagenase hydrolyzed native and denatured collagen and synthetic peptides with sequences of 2,4-dinitrophenyl-l-prolyl-l-leucylglycyl-l-isoleucyl-l-alanylglycyl-l-arginine amide and 2,4-dinitrophenyl-l-prolyl-l-glytaminyl-l-glycyl-l-isoleucyl-l-alanylglycyl-l-glutaminyl-d-arginine. The specific enzyme activity against the latter substrate and collagen fibrils is found to be 933 nmol/min per mg protein and 8440 units (μg collgen degraded/min), respectively. Optimum pH for the enzyme is 7.5-8.5. A collagenase complex with α2-macroglobulin did not hydrolyze collage fibrils, but digested the synthetic substrates at the Gly-Ile bond. The amino acid composition of the enzyme was determined. Immunoelectrophoresis of the enzyme at pH 8.6 against anti-tadpole collagenase rabbit immunoglobulin G shows a single precipitin line at a position migrating faster than human serum albumin and corresponding to enzyme activity against collagen fibril and synthetic substrates. © 1979.