METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN

结缔组织基质分解中的金属蛋白酶

• 批准号: 2694411
• 负责人: HIDEAKI NAGASE
• 金额: $ 22.22万
• 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/2694411
• 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.

基质金属蛋白水解酶(MMPs)是一组蛋白水解酶 对细胞外基质的降解至关重要。他们表演的是 哺乳动物生物体和疾病中的各种重要功能 如关节炎牙周炎、动脉粥样硬化和肿瘤细胞 侵袭和转移。已经确定了17个MMP，包括 四个胶原酶，两个明胶酶，两个基质蛋白，四个膜型 MMPs(MT-MMPs)，以及其他五种。首席调查员已经为许多人 多年来为我们对基质金属蛋白酶的理解做出了重大贡献 函数，并且在此应用程序中，他请求支持以在 有两个方面。 在第一个具体目标中，首席调查员建议定义 胶原酶结构的那些方面使其具有解离能力 从而裂解胶原蛋白的三股螺旋。这将使用以下工具完成 产生嵌合酶或突变酶的重组技术。 基质金属蛋白酶-1和基质金属蛋白酶-1/基质金属蛋白酶-3嵌合体与合成三联体的相互作用 模拟序列的Helica多肽(由合作者Dr.Fields提供) I型、III型和III型胶原的裂解部位周围 也将使用荧光偏振和微量热法进行检查 技巧。此外，与校长博德博士合作 研究人员的目标是确定催化的晶体结构 失活的基质金属蛋白酶-1与一个或多个三螺旋多肽形成的复合体。 第二个特定的目的是解决原基质金属蛋白酶2在细胞中的激活 表面，这一过程似乎对癌细胞的转移至关重要。一个 基质金属蛋白酶原2活性的定量检测将使用 放射性标记的催化失活的前基质金属蛋白酶2.细胞的重要性 MT1-基质金属蛋白酶分子在活化过程中的附着和结构 基质金属蛋白酶-将通过构建和细胞表面表达来检测 分别含有催化端和C端的MMP1/MT1-MMP型嵌合体 MMP1的结构域。首席研究员还建议研究 细胞表面TIMP-2在这一激活过程中的作用。