METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN

结缔组织基质分解中的金属蛋白酶

• 批准号: 6374903
• 负责人: HIDEAKI NAGASE
• 金额: $ 20.42万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374903
• 关键词: Mammalia; antisense nucleic acid; cell adhesion; chimeric proteins; collagenase; connective tissue metabolism; enzyme activity; enzyme induction /repression; enzyme structure; extracellular matrix; fluorescence polarization; human tissue; metalloendopeptidases; microcalorimetry; protein folding; protein structure function; stromelysin; tissue /cell culture; tissue inhibitor of metalloproteinases; zymogens

The matrix metalloptroteinases (MMPs) are a group of proteinases that are central to the degradation of extracellular matrix. They perform a variety of critical functions in the mammalian organism and in diseases such as arthritis periodontitis, atherosclerosis, and in tumor cell invasion and metastasis. Seventeen MMPs have been identified including four collagenases, two gelatinases, two stromelysins, four membrane-type MMPs (MT-MMPs), and five others. The principal investigator has for many years been making major contributions to our understanding of MMP function, and in this application he requests support to continue this in two areas. In the first specific aim the principal investigator proposes to define those aspects of collagenase structure that give it the ability to unwind and thus cleave the collagen triple helix. This will be done using recombinant technologies to generate chimeric or mutant enzymes. The interaction of MMP-1 and MMP-1/MMP-3 chimeras with synthetic triple helica peptides (supplied by collaborator Dr. Fields) that mimic sequences around the cleavage site of collagens with type I, II and III collagens will also be examined using fluorescent polarization and microcalorimetry techniques. In addition in collaboration with Dr. Bode the principal investigator aims to determine the crystal structure of catalytically inactive MMP-1 in complex wit one or more triple helical peptides. The second specific aim addresses the activation of proMMP 2 at the cell surface, a process that appears critical for cancer cell metastasis. A quantitative assay for proMMP 2 activation will be developed using radiolabelled catalytically inactive proMMP 2. The importance of cell attachment and the structure of the MT1-MMP molecule in the activation of MMP-will be examined by construction and cell surface expression of MMP-1/MT1-MMP chimeras containing separately the catalytic and C-terminal domains of MMP 1. The principal investigator also proposes to examine the role of cell surface TIMP-2 in this activation process.

基质金属蛋白酶（MMP）是一组蛋白酶， 细胞外基质降解的中心。 它们执行 哺乳动物机体和疾病中的各种关键功能 如关节炎、牙周炎、动脉粥样硬化，以及肿瘤细胞中 侵袭和转移。 已经确定了17种MMP，包括 四种胶原酶，两种明胶酶，两种溶基质素，四种膜型 MMPs（MT-MMPs）和其他5种。 首席研究员对许多 多年来为我们理解MMP做出了重大贡献 功能，并在此应用程序中，他请求支持继续这一点， 两个领域。 在第一个具体目标中，主要研究者建议定义 胶原酶结构的那些方面， 从而切割胶原三螺旋。 这将使用 重组技术以产生嵌合或突变酶。 MMP-1和MMP-1/MMP-3嵌合体与合成三联体的相互作用 模拟序列的螺旋肽（由合作者Fields博士提供） I型、II型和III型胶原的胶原裂解位点周围 还将使用荧光偏振和微量热法进行检查 技术. 此外，在与博德博士的合作， 研究人员旨在确定催化剂的晶体结构， 与一个或多个三螺旋肽复合失活MMP-1。 第二个具体目标是在细胞中激活proMMP 2 表面，这是一个对癌细胞转移至关重要的过程。 一 将开发用于proMMP 2活化的定量测定， 放射性标记的无催化活性的proMMP 2。 细胞的重要性 MT 1-MMP分子的结构和粘附在激活 MMP-2将通过构建和细胞表面表达来检查， MMP-1/MT 1-MMP嵌合体分别含有催化和C-末端 MMP 1的结构域。 首席研究员还建议审查 细胞表面TIMP-2在此活化过程中的作用。