DeClerck Y A
Division of Hematology-Oncology, Childrens Hospital, Los Angeles, California 90027.
Arch Biochem Biophys. 1988 Aug 15;265(1):28-37. doi: 10.1016/0003-9861(88)90367-0.
A potent polypeptide inhibitor of mammalian collagenases was purified to homogeneity from medium conditioned by bovine aortic smooth muscle cells maintained in culture. This inhibitor was purified by a series of molecular sieve and heparin-Sepharose chromatographic procedures; it had an apparent Mr of 28,500 and was a major protein secreted by the smooth muscle cells. It was found to be active against several mammalian collagenases including those obtained from rabbit and human fibroblasts and a tumor-specific type IV collagenase. In contrast, it had minimal inhibitory activity for bacterial collagenase and was inactive against the serine proteases plasmin and trypsin. The inhibitor shared many characteristics with tissue inhibitor of metalloproteinases including the ability to irreversibly inhibit susceptible proteinases, heat and acid resistance, and sensitivity to trypsin degradation and reduction-alkylation. A polyclonal rabbit antiserum with blocking activity which recognized the Mr 28,500 protein was obtained. This inhibitor, which is likely produced by bovine vascular smooth muscle cells in vivo to protect the collagen matrix of blood vessels, may play an important role in pathological conditions associated with alteration of collagen metabolism in tissues.
一种有效的哺乳动物胶原酶多肽抑制剂，从培养的牛主动脉平滑肌细胞条件培养基中纯化至同质。该抑制剂通过一系列分子筛和肝素 - 琼脂糖色谱法进行纯化；其表观分子量为28,500，是平滑肌细胞分泌的主要蛋白质。发现它对几种哺乳动物胶原酶具有活性，包括从兔和人成纤维细胞获得的胶原酶以及肿瘤特异性IV型胶原酶。相比之下，它对细菌胶原酶的抑制活性最小，对丝氨酸蛋白酶纤溶酶和胰蛋白酶无活性。该抑制剂与金属蛋白酶组织抑制剂具有许多共同特征，包括不可逆抑制敏感蛋白酶的能力、耐热性和耐酸性，以及对胰蛋白酶降解和还原烷基化的敏感性。获得了一种具有阻断活性的多克隆兔抗血清，它能识别分子量为28,500的蛋白质。这种抑制剂可能由牛血管平滑肌细胞在体内产生，以保护血管的胶原基质，可能在与组织中胶原代谢改变相关的病理状况中起重要作用。
