PROTEOGLYCAN STRUCTURE, FUNCTION AND METABOLISM

蛋白多糖是结缔组织中普遍存在的成分。他们经常 作为这些组织细胞外基质的主要成分出现。 软骨蛋白多糖一直是研究最彻底的，他们已经 作为研究蛋白多糖结构和功能的有用模型 在其他更复杂的结缔组织基质中。详细分析了 蛋白多糖结构通过传统的生化程序已经被 由于其固有的异质性和多分散性， 蛋白多糖本身。免疫学方法提供了一种 产生特异性识别不同抗原的抗体 存在于蛋白多糖和链接蛋白上的决定因素。最近的 单抗程序的引入提供了这种可能性。 产生特异性识别结构和结构的抗体 生物大分子上的功能性抗原决定簇。在这 我们计划生产和鉴定单克隆抗体的提案 识别蛋白多糖和连接蛋白上的特定决定因素。这个 抗体的特异性将使用生化方法来确定， 以及在免疫分析程序中。特定的抗体将应用于 组织的定量、鉴定、提纯和测定问题 蛋白多糖和连接蛋白的分布。两项免疫化学研究 将采用隔离材料和免疫组织化学方法。 这项提议的一个主要目标是使用特定的单抗 作为鉴定和纯化LINK多肽片段的手段 蛋白质和蛋白多糖。这些多肽将用于测定 连接蛋白的氨基酸序列(一级结构)和 蛋白多糖。此外，识别细胞功能方面的抗体 蛋白多糖和连接蛋白的相互作用将用于研究 确定与蛋白多糖有关的结构功能关系 集合体的形成。更长期的目标是将这些研究扩展到 更好地了解蛋白多糖在其他方面的代谢 复杂的结缔组织。这些研究将增加我们对 结缔组织在正常情况下和疾病中起作用。

Proteoglycans are ubiquitous components of connective tissues. They often occur as major components of the extra-cellular matrix of these tissues. Cartilage proteoglycans have been the most thoroughly studied and they have served as a useful model for examining proteoglycan structure and function in other more complex connective tissue matrices. Detailed analysis of proteoglycan structure by conventional biochemical procedures has been complicated by the inherent heterogeneity and polydisperisty of the proteoglycans themselves. Immunological methods offer the possibililty of raising antibodies which specifically recognize different antigenic determinants present on proteoglycans and link protein. The recent introduction of monoclonal antibody procedures has offered the possiblity of producing antibodies which specifically recognize structural and functional antigenic determinants on biological macromolecules. In this proposal we plan to produce and characterize monoclonl antibodies that recognize specific determinants on proteoglycan and link protein. The specificity of the antibodies will be determined using biochemical methods, and in immunoassay procedures. Specific antibodies will be applied to problems of quantitating, identifying, purifying and determining the tissue distribution of proteoglycan and link protein. Both immunochemical studies on isolated material and immunohistochemical procedures will be employed. A major objective of this proposal is to use specific monoclonal antibodies as a means of characterizing and purifying peptide fragments of link protein and proteoglycan. These peptides will be used in the determination of the amino acid sequences (primary structure) of link protein and lproteoglycan. In addition, antibodies recognizing functional aspects of proteoglycan and link protein interactions will be used instudies determining structure function relationships involved in proteoglycan aggregate formation. A longer term objective is to extend these studies to gain a better understanding of proteoglycan metabolism in other more complex connective tissues. These studies will increase our knowledge of connective tissue function in normal conditions and in disease.