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-arginie amide and 2,4-dinitrophenyl-L-prolyl-L-glutaminyl-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 (microgram collagen degraded/min), respectively. Optimum pH for the enzyme is 7.5-8.5. A collagenase complex with alpha2-macroglobulin did not hydrolyze collagen 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.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies; D001001:Anura; D003094:Collagen; D003012:Microbial Collagenase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D010455:Peptides; D011892:Rana catesbeiana / Q000378:metabolism*; D013379:Substrate Specificity; D000511:alpha-Macroglobulins / Q000494:pharmacology
D000596:Amino Acids; D000906:Antibodies; D010455:Peptides; D000511:alpha-Macroglobulins; D003094:Collagen; D003012:Microbial Collagenase