PROTEOGLYCAN STRUCTURE FUNCTION AND METABOLISM

蛋白聚糖是结缔组织中普遍存在的成分， 作为这些组织的细胞外基质的主要成分发生。 鲤鱼蛋白聚糖已被最彻底地研究， 它们作为了解蛋白聚糖结构的有用模型 并在其他更复杂的结缔组织基质中发挥作用。 最近 利用重组DNA技术取得的进展， 推进我们的知识，以初级氨基酸序列数据的核心 这些结缔组织蛋白聚糖的大量蛋白质。 这些 研究使研究人员能够将几类 蛋白聚糖分为基因和结构/功能相关家族。 是 现在可以使用这些新开发的分子生物学技术 进行各种各样的细胞生物学和代谢研究 研究了几个迄今为止无法解决的与蛋白聚糖有关的问题， 结构、功能和代谢。 尽管取得了这些进展，我们仍然对这方面知之甚少。 固有异质性和多分散性的生物学意义 这在不同组织中发现的各种蛋白聚糖中观察到， 结缔组织 这种异质性很大程度上是一种复杂的 在这些分子中发生的一系列翻译后修饰 在它们的生物合成过程中，在它们的组织和周转过程中， 在细胞外基质中。 目前，免疫学方法提供了 这可能是研究导致癌症的生物学机制的最好方法。 在体内观察到固有的蛋白聚糖异质性。 过去十 多年来，该实验室在单克隆抗体的使用领域处于领先地位。 抗体技术研究蛋白多糖的结构、功能和 健康和疾病的代谢。 在过去的一段时间里， 并鉴定了几种新的单克隆抗体， 识别细微结构成分的差异表达， 结缔组织蛋白多糖在生长和发育过程中， 病理学，例如骨关节炎。 在目前的提案中，我们计划 继续开发和使用单克隆抗体技术， 结缔组织中蛋白多糖的结构和功能 组织中 这项建议的重点将是蛋白多糖的异质性， 软骨 为达致这个目标，我们的具体目标是：（i）进一步 表征目前存在的抗蛋白聚糖的单克隆抗体 表位;（ii）产生一组新的单克隆抗体， 针对域特异性、种特异性和组织特异性 不同蛋白聚糖亚群上的表位;（iii）产生 针对蛋白聚糖上的“新表位”的单克隆抗体， 由于这些分子的金属蛋白酶降解而发生， (iv)使用在前三个特定目标中产生的抗体， 研究蛋白多糖异质性的机制。

Proteoglycans are ubiquitous components of connective tissues where they occur as major components of the extracellular matrix of these tissues. Cartilage proteoglycans have been the most thoroughly studied and as such they have served as a useful model for understanding proteoglycan structure and function in other more complex connective tissue matrices. Recent advances made the use of recombinant DNA technologies have significantly advanced our knowledge as to primary amino acid sequence data of the core proteins of a large number of these connective tissue proteoglycans. These studies have allowed researchers to categorize several classes of the proteoglycans into gene- and structure/function-related families. It is now possible to use these newly developed molecular biological technologies to perform a wide variety of cell biological and metabolic studies examining several hitherto unattainable problems relating to proteoglycan structure function and metabolism. In spite of these advances, we still know very little regarding the biological significance of the inherent heterogeneity and polydispersity that is observed in the wide variety of proteoglycans found in different connective tissues. This heterogeneity is largely a result of a complex array of post-translational modifications that occur in these molecules during both their biosynthesis, and also in their organization and turnover within the extracellular matrix. At present immunological approaches offer possibly the best means of studying the biological mechanism causing the inherent proteoglycans heterogeneity observed in vivo. Over the past ten years this laboratory has pioneered the field in the use of monoclonal antibody technology to study proteoglycan structure, function and metabolism in health and disease. In the past funding period we produced and characterized several new monoclonal antibodies that were used to identity the differential expression of subtle structural components in connective tissue proteoglycans during growth and development and in pathology, e.g. osteoarthritis. In this current proposal we plan to continue to develop and use monoclonal antibody technology to address problems relating to proteoglycan structure and function in connective tissues. The focus of this proposal will be proteoglycan heterogeneity in cartilage. Our specific aims to achieve this objective are (i) to further characterize currently existing monoclonal antibodies against proteoglycan epitopes; (ii) to produce a panel of new monoclonal antibodies with specificities directed against domain-, species- and tissue-specific epitopes on different proteoglycan subpopulations; (iii) to produce monoclonal antibodies directed against "neo-epitopes" on proteoglycans that occur as a result of metalloproteinase degradation of these molecules, and (iv) to use the antibodies generated in the previous three specific aims to study mechanisms involved in proteoglycan heterogeneity.