Hori H, Nagai Y
Biochim Biophys Acta. 1979 Jan 12;566(1):211-21. doi: 10.1016/0005-2744(79)90263-8.
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.
蝌蚪胶原酶可水解天然和变性胶原以及具有2,4 - 二硝基苯基 - L - 脯氨酰 - L - 亮氨酰甘氨酰 - L - 异亮氨酰 - L - 丙氨酰甘氨酰 - L - 精氨酸酰胺和2,4 - 二硝基苯基 - L - 脯氨酰 - L - 谷氨酰胺基 - 甘氨酰 - L - 异亮氨酰 - L - 丙氨酰甘氨酰 - L - 谷氨酰胺基 - D - 精氨酸序列的合成肽。发现该酶对后一种底物和胶原纤维的比酶活性分别为每毫克蛋白质933 nmol/分钟和8440单位（每分钟降解的微克胶原）。该酶的最适pH为7.5 - 8.5。与α2 - 巨球蛋白形成的胶原酶复合物不水解胶原纤维，但在甘氨酰 - 异亮氨酸键处消化合成底物。测定了该酶的氨基酸组成。在pH 8.6条件下，用抗蝌蚪胶原酶兔免疫球蛋白G对该酶进行免疫电泳，结果显示在比人血清白蛋白迁移速度快的位置出现一条单一沉淀线，且该位置与针对胶原纤维和合成底物的酶活性相对应。
