PURIFICATION OF PROCOLLAGENASE AND COLLAGENASE BY AFFINITY CHROMATOGRAPHY ON SEPHAROSE-COLLAGEN
PURIFICATION OF PROCOLLAGENASE AND COLLAGENASE BY AFFINITY CHROMATOGRAPHY ON SEPHAROSE-COLLAGEN
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DOI：
10.1016/0014-5793(77)80768-0
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发表时间：
1977-01-01
期刊：
影响因子：
3.5
通讯作者：
VAES, G
中科院分区：
文献类型：
--
作者：
GILLET, C;EECKHOUT, Y;VAES, G
特定动物胶原酶（EC 3.4）。24.3)已被几种方法纯化，包括亲和层析的胶原偶联琼脂糖[l-3]或包埋在聚丙烯酰胺珠[4]。潜伏形式的胶原酶或前胶原酶的发生目前已被充分证实，但它们的分子结构——酶原或酶抑制剂复合物——及其在生理或病理中的作用仍有待阐明。本报告表明，培养[6]的小鼠骨外植体释放的前胶原酶对偶联琼脂糖的天然盐溶性胶原具有显著的亲和力，并且可以通过亲和层析广泛纯化（比活性约为3500单位/毫克蛋白质）。该方法高效、快速、简便，在其它哺乳动物（包括人）前胶原酶的纯化中应具有广泛的适用性。这项工作的初步摘要已于2010年提出。
Specific animal collagenases (EC 3.4. 24.3) have been purified by several methods including affinity chromatography on collagen coupled to agarose [l-3] or entrapped in polyacrylamide beads [4]. The occurrence of latent forms of collagenase-or procollagenases-is now well documented [5] but their molecular structure-zymogens or enzyme-inhibitor complexes-and their role in physiology or pathology remain to be elucidated. The present report demonstrates that the procollagenase which is released by mouse bone explants in culture [6] exhibits a striking affinity for native salt-soluble collagen coupled to agarose and that it can be extensively purified (to a specific activity of about 3500 units/mg protein) by affinity chromatography. This efficient, rapid and simple method should have broad applicability in the purification of other mammalian (including human) procollagenases. A preliminary abstracted account of this work has already been presented [7].