Evidence for the presence of a latent form of collagenase in human rheumatoid synovial fluid.
Evidence for the presence of a latent form of collagenase in human rheumatoid synovial fluid.
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人类类风湿滑液中存在潜在形式的胶原酶的证据。
DOI：
10.1093/oxfordjournals.jbchem.a129846
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发表时间：
1972
影响因子：
2.7
通讯作者：
Y. Nagai
中科院分区：
文献类型：
--
作者：
S. Abe;Y. Nagai
众所周知，血清 a-球蛋白通常会抑制组织蛋白酶 (1)，可能作为维持组织中蛋白水解活性生理水平的控制因素 (2)。动物胶原酶*是一种高特异性组织蛋白酶，可裂解天然和变性胶原蛋白，在生理条件下在胶原蛋白代谢中发挥作用。最近艾森等人。据报道，动物和人类胶原酶受到血清中的 a2-巨球蛋白 (a2M) 和抗胰蛋白酶的抑制 (3)，并证明新鲜人皮肤提取物中存在免疫反应性但酶活性失活的胶原酶，表明提取物中的这些抑制剂掩盖了酶活性 (4)。 Nagai 和 Hori (5) 发现，在酶与未鉴定的组织成分之间存在可逆相互作用的选定条件下，可以直接从脊椎动物皮肤和滑膜匀浆中分离出活性形式的胶原酶。这些事实促使我们研究酶和 a2M 之间的相互作用，特别参考球蛋白在胶原蛋白代谢中可能的生理功能。由此表明，纯化的蝌蚪胶原酶与天然a2M紧密结合，形成无酶活性的复合物，该复合物在凝胶过滤时未分离地被洗脱。在用于解离胶原酶-胶原复合物(pH 5.0)(6)或抗原-抗体复合物(pH 2.5)的条件下，胶原酶没有与复合物分离。然而，最近开发的用于解离多种抗原抗体复合物 (7, 8) 的 0.015 M 磷酸盐缓冲液（pH 6.0）中的 3 M 硫氰酸钠，被发现可通过选择性地使 a2M 变性，有效地从无活性酶 -a2M 复合物中释放活性形式的胶原酶。** 在本通讯中，我们报告说，该试剂在应用于人类类风湿滑液时，引起了激活体液中非活性（潜在）形式的胶原酶。这一发现为人类类风湿滑液中存在潜在形式的胶原酶提供了证据。还讨论了潜在酶在该疾病中的发病机制。
It is well known that serum a-globulins generally inhibit tissue proteases (1), possibly acting as controlling factors in maintaining physiological levels of the proteolytic activi ties in tissues (2). Animal collagenase,* a tissue protease of high specificity which cleaves native and denatured collagen, is supposed to play a role in collagen metabolism under physiological conditions. Recently Eisen et al. reported that animal and human collagenases were inhibited by a2-macroglobulin (a2M) and atantitrypsin in serum (3) and demonstrated the presence of immunoreactive, but enzymatically inactive, collagenase in fresh human skin extracts, suggesting that the enzyme activity is masked by these inhibitors in the extracts (4). Nagai and Hori (5) found that collagenases could be directly isolated in an active form from vertebrate skin and synovial membrane homogenates under selected conditions where a reversible interaction between the enzyme and unidentified tissue components existed. These facts prompted us to study the interaction between the enzyme and a2M with special references to the possible physiological function of the globulin in collagen metab olism. It was thus shown that purified tadpole collagenase combined tightly with native a2M, forming a enzymatically inactive complex which was eluted unseparated on gel filtration. The collagenase was not separated from the complex under conditions employed for the dissociation of either collagenase-col lagen complex (pH 5.0)(6) or antigen-antibody complex (pH 2.5). However, 3 M sodium thiocyanate in 0.015 M phosphate buffer, pH 6.0, recently developed for the dissociation of a number of antigen-antibody complexes (7, 8), was found to be effective in releasing the collagenase in an active form from the inac tive enzyme-a2M complex by selectively de naturing a2M.** In this communication, we report that this reagent, when applied to human rheumatoid synovial fluids, caused an activation of an inactive (latent) form of col lagenase in the fluids. This finding provides evidence for the presence of a latent form of collagenase in human rheumatoid synovial fluids. Pathogenetic roles of the latent en zvme in the disease are also discussed.