PROTEOGLYCAN STRUCTURE FUNCTION AND METABOLISM

蛋白多糖是结缔组织中普遍存在的成分，在这些组织中 作为这些组织细胞外基质的主要成分而发生。 软骨蛋白多糖一直是研究最彻底的，因此 它们已成为了解蛋白多糖结构的有用模型。 并在其他更复杂的结缔组织基质中发挥作用。近期 利用重组DNA技术取得的进展显著地 增进了我们对岩心初级氨基酸序列数据的了解 大量这种结缔组织蛋白多糖的蛋白质。这些 研究已经使研究人员能够将几类 蛋白多糖进入与基因和结构/功能相关的家族。它是 现在有可能使用这些新开发的分子生物技术 进行广泛的细胞生物学和代谢研究 研究与蛋白多糖有关的几个迄今无法解决的问题 结构、功能和新陈代谢。 尽管取得了这些进展，但我们仍然对 天然异质性和多分散性的生物学意义 在不同种类的蛋白多糖中可以观察到这一点 结缔组织。这种异质性在很大程度上是复杂的 在这些分子中发生的一系列翻译后修饰 在它们的生物合成和组织和周转过程中 在细胞外基质内。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 体内观察到固有的蛋白多糖异质性。在过去十年中 多年来，这个实验室在使用克隆方面一直是该领域的先驱 抗体技术用于研究蛋白多糖的结构、功能和 健康和疾病中的代谢。在过去的融资期间，我们生产了 并鉴定了几种新的用于 中微妙结构成分的差异化表达 结缔组织蛋白多糖在生长发育中的作用 病理学，例如骨性关节炎。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来解决 连接性蛋白多糖结构和功能的相关问题 纸巾。这项建议的重点将是蛋白多糖的异质性 软骨。我们实现这一目标的具体目标是：(I)进一步 鉴定目前已有的抗蛋白多糖的单抗 表位；(Ii)产生一组新的单抗 针对特定领域、特定物种和特定组织的特异性 不同蛋白多糖亚群上的表位；(Iii)产生 针对蛋白多糖上的“新表位”的单抗 发生于金属蛋白酶对这些分子的降解，以及 (四)利用前三个特定目标产生的抗体 研究与蛋白多糖异质性有关的机制。

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.