ROLE OF COLLAGENOLYTIC METALLOPROTEINASES IN METASTASES

为探讨基质金属蛋白酶(MMPs)在心肌梗死中的作用。 肿瘤的侵袭和转移，我们已经关注了多个层面 对这些酶的调节。研究表明，与 基质金属蛋白酶家族的其他成员，72 kDa明胶酶A水平为 增加对TGFbeta1的反应，不受促癌作用的影响 佛波酯，并显示结直肠、乳房、甲状腺、 卵巢和膀胱癌组织与邻近正常组织的比较 粘膜组织。我们已经确定了一种细胞激活机制， 细胞表面是否与72 kDa明胶酶A相关且具有特异性 酶，可通过佛波酯或佛波醇酯预处理而诱导 刀豆蛋白A这种细胞激活机制不影响其他 胶原酶基因家族的成员。此激活机制将出现 要求细胞表面结合明胶酶A酶，我们有 鉴定出一种可能的明胶酶A受体。 我们对潜伏酶TIMP-2复合体的结构进行了研究 酶缺失突变体的产生及酶抑制剂的交联化 学习。这些研究表明，72 kDa的明胶酶A在 至少两个TIMP-2结合域。主结合结构域被定位 在C-末端，酶的类血凝蛋白结构域。这个结合部位 以潜伏酶的形式存在。第二个结合部位在 酶活性部位，只有在有机汞之后才可用 介导酶的激活。 最后，92 kDa明胶酶B的抗肽抗体， 间质胶原酶、基质分解素-1和基质分解素-2已被 准备好了并进行了表征。

In order to investigate the role of matrix metalloproteinases (MMP) in tumor invasion and metastases, we have focused on the multilevel regulation of these enzymes. Studies have shown that in contrast with other members of the MMP enzyme family, 72 kDa gelatinase A levels are increased in response to TGFbeta1, are unaffected by the tumor promoting phorbol esters, and show elevated levels in colorectal, breast, thyroid, ovarian and bladder tumor tissues when compared with adjacent normal mucosa tissues. We have identified a cellular activation mechanism which is cell surface associated and specific for the 72 kDa gelatinase A enzyme, and which can be induced by pretreatment with phorbol esters or concanavalin A. This cellular activation mechanism does not affect other members of the collagenase gene family. This activation mechanism appears to require cell surface binding of the gelatinase A enzyme, and we have identified a putative gelatinase A receptor. We have studied the structure of the latent enzyme TIMP-2 complex through production of enzyme deletion mutants and enzyme inhibitor cross linking studies. These studies demonstrate that the 72 kDa gelatinase A has at least two TIMP-2 binding domains. The principal binding domain is located in the C-terminal, hemopexin-like domain of the enzyme. This binding site is available in the latent enzyme form. The second binding site is at the enzyme active site and only becomes available following organomercurial mediated enzyme activation. Finally, antipeptide antibodies against the 92 kDa gelatinase B, interstitial collagenase, stromelysin-1 and stromelysin-2 have been prepared and characterized.