NOVEL METALLOPROTEINASE INHIBITORS--ROLE IN TUMOR INVASION AND METASTASIS

新型金属蛋白酶抑制剂——在肿瘤侵袭和转移中的作用

• 批准号: 3774378
• 负责人: W G STETLER-STEVENSON
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3774378
• 关键词: X ray crystallography; angiogenesis; basement membrane; blood brain barrier; cell cycle; clone cells; collagenase; enzyme complex; enzyme mechanism; enzyme structure; genetic transcription; growth inhibitors; human tissue; membrane reconstitution /synthesis; metastasis; neoplasm /cancer invasiveness; nuclear magnetic resonance spectroscopy; protein sequence; proteolysis; stroke; structural genes; synthetic peptide; tissue inhibitor of metalloproteinases; vascular endothelium

We have isolated and characterized the complete primary structure of a new member of the tissue inhibitor of metalloproteinase family (TIMP family) which we refer to as TIMP-2. Recent studies have shown that all cells studied to date which secrete the 72 kDa type IV collagenase enzyme secrete this enzyme as a complex with TIMP-2. The majority greater than 95% of secreted 72 kDa gelatinase is found in complexed form as most cells tested secrete 2-4 fold as free TIMP-2. Our studies have shown that TIMP- 2 transcript ion is regulated independently of both TIMP-1 and the 72 kDa type IV collagenase enzyme. We have also demonstrated that TIMP-2 is anti-angiogenic. The mechanism for this effect is two fold; through inhibition of endothelial cell proliferation and blocking endothelial cell-mediated matrix proteolysis. TIMP-2 inhibits tumor cell invasion through reconstituted basement membranes in vitro, and this inhibitor demonstrates erythroid potentiating activity (EPA) . TIMP-2 inhibits proteolytic opening of the blood brain barrier in hemorrhagic stroke models. TIMP-2 genomic clones have been obtained and partial sequencing has identified two introns in the 3' end of the gene. The gene appears to be single copy and is localized on human chromosome 17q25. We have examined the TIMP-2 protein structure and have localized the metalloprotease inhibitory domain to the N-terminal half of the molecule. Further sublocalization has been attempted using a synthetic peptide approach as well as protein crystallization for x-ray diffraction and NMR- spectroscopy.

我们已经分离并表征了一种新的完整的初级结构 金属蛋白酶家族组织抑制剂（TIMP家族）成员 我们将其称为 TIMP-2。 最近的研究表明，所有细胞 迄今为止研究的分泌 72 kDa IV 型胶原酶的酶 分泌该酶与 TIMP-2 的复合物。 大多数大于 与大多数细胞一样，95% 的分泌型 72 kDa 明胶酶以复合形式存在 经测试，其分泌量是游离 TIMP-2 的 2-4 倍。 我们的研究表明 TIMP- 2 转录离子的调节独立于 TIMP-1 和 72 kDa IV 型胶原酶。 我们还证明了 TIMP-2 抗血管生成。 这种效应的机制有两个方面：通过 抑制内皮细胞增殖并阻断内皮细胞 细胞介导的基质蛋白水解。 TIMP-2抑制肿瘤细胞侵袭 通过体外重建基底膜，并且该抑制剂 表现出红细胞增强活性（EPA）。 TIMP-2 抑制 出血性中风血脑屏障的蛋白水解开放 模型。 TIMP-2基因组克隆已获得，部分测序已完成 鉴定出该基因 3' 端有两个内含子。 该基因似乎是 单拷贝，位于人类染色体 17q25 上。 我们检查了 TIMP-2 蛋白质结构并定位了 金属蛋白酶抑制结构域连接到分子的 N 端一半。 使用合成肽尝试进一步亚定位 方法以及用于 X 射线衍射和 NMR 的蛋白质结晶 光谱学。