STRUCTURE/FUNCTION OF METALLOPROTEINASE MT3/MMP

金属蛋白酶 MT3/MMP 的结构/功能

• 批准号: 2450610
• 负责人: DUANQING PEI
• 金额: $ 13.78万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-05 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2450610
• 关键词: enzyme structure; extracellular matrix; gene expression; metalloendopeptidases; metastasis; neoplastic process; polymerase chain reaction; protein structure function; proteolysis; recombinant proteins; zymogens

The long term objective of this proposal is to understand the proteolytic machinery employed by malignant tumor cells to traverse the dense structural barriers established by the extracellular matrix during the invasive and metastatic process. Despite improved surgical techniques against the primary tumors, failure to control the spread of cancer cells contributes to most of the cancer related death today. Cancer cells invade local tissues and metastasize to distant organs by regulating the expression of proteolytic enzymes that allow them to degrade their surrounding extracellular matrix barriers. One family of proteinases, known as the matrix-degrading metalloproteinases, have been implicated in cancer progression by virtue of their ability to degrade all proteinaceous components of the extracellular matrix including collagens, elastin and proteoglycans. While the majority of the MMPs are secreted enzymes, the recently identified MT-MMP subgroup contain potential transmembrane domains at their C-termini and therefore be able to anchor on the cell surface and form a localized proteolytic zone between the invading cells and their surrounding ECM barrier. The discovery of this subgroup clarified earlier suspicion that tumor cells utilize special proteolytic machinery anchored on their cell surfaces to clear a pathway during local invasion and metastasis. MT3-MMP, a recent addition to this subgroup, was recently identified from an malignant oral melanoma tissue by degeneraate RT-PCR strategy and found to be expressed by other cells and tissues. Furthermore, it has been proposed to play an important role in the development of invasive phenotype of tumor. To establish a causal relationship between MT3-MMP expression and the invasive phenotype, a comprehensive strategy is proposed in this application to 1) elucidate the mechanism(s) governing its conversion from zymogen to active form, 2) characterize its enzymic properties by expressing and isolating the enzyme in recombinant forms, and 3) assess the ability of MT3-MMP to regulate the invasive potential of tumor cells in an in vitro construct of the extracellular matrix. Insights from the characterization of MT3-MMP proteolytic activity, its activation mechanism as well as its ability to confer invasive potential on tumor cells should not only provide a thorough knowledge base on the role of this cell membrane associated metalloproteinase in human malignancies, but also its potential utility as a diagnotic marker or a target for new cancer therapies.

该提案的长期目标是了解 恶性肿瘤细胞利用蛋白水解机制来遍历 细胞外基质在过程中建立的致密结构屏障 侵袭和转移过程。尽管手术有所改善 针对原发肿瘤的技术，未能控制扩散 当今大多数与癌症相关的死亡都是由癌细胞造成的。 癌细胞通过以下途径侵入局部组织并转移至远处器官 调节蛋白水解酶的表达，使它们能够 降解其周围的细胞外基质屏障。 一个家庭 蛋白酶，又称基质降解金属蛋白酶，已被 由于其降解能力而与癌症进展有关 细胞外基质的所有蛋白质成分，包括 胶原蛋白、弹性蛋白和蛋白聚糖。 虽然大多数 MMP 是分泌酶，最近鉴定的 MT-MMP 亚组包含 潜在的跨膜结构域位于其 C 端，因此能够 锚定在细胞表面并形成局部蛋白水解区 入侵细胞与其周围的 ECM 屏障之间。 这 这一亚组的发现澄清了先前的怀疑：肿瘤细胞 利用锚定在细胞表面的特殊蛋白水解机制 在局部侵袭和转移过程中清除通路。 MT3-MMP，一个 最近添加到这个亚组，最近从一个 通过退化RT-PCR策略发现恶性口腔黑色素瘤组织 由其他细胞和组织表达。 此外，已经 提议在侵入性技术的发展中发挥重要作用 肿瘤的表型。 建立MT3-MMP之间的因果关系 表达和侵袭表型，综合策略是 本申请中提出的目的是 1) 阐明控制机制 其从酶原到活性形式的转化，2) 表征其酶活性 通过以重组形式表达和分离酶来确定特性， 3）评估MT3-MMP调节侵袭潜力的能力 细胞外基质的体外构建体中的肿瘤细胞。 MT3-MMP 蛋白水解活性表征的见解 激活机制及其赋予侵入潜力的能力 关于肿瘤细胞的研究不仅应该提供关于肿瘤细胞的全面知识基础 这种细胞膜相关金属蛋白酶在人体中的作用 恶性肿瘤，而且其作为诊断标志物的潜在用途或 新癌症疗法的目标。