BIOLOGY AND CHEMISTRY OF PROCOLLAGEN IN CELL CULTURE

细胞培养中原胶原的生物学和化学

• 批准号: 3154938
• 负责人: MARVIN L TANZER
• 金额: $ 13.81万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-09-01 至 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3154938
• 关键词: carbohydrate structure; chondrocytes; chondroitin sulfates; collagen disorder; complementary DNA; connective tissue metabolism; fibroblasts; genetic library; genetic regulation; genome; human tissue; immunoelectron microscopy; molecular cloning; molecular pathology; mutant; oligosaccharides; orphan disease /drug; proteoglycan; radiotracer; sulfation; tissue /cell culture

This is a competing continuation request for another five years of research support. The major focus of the proposed studies is upon the structure, function and biological expression of macromolecules of the extracellular matrix, examining the protein and carbohydrate structure of laminin and the gene structure for the core protein of chondroitin sulfate proteoglycan. Laminin is isolated from the EHS tumor of mice and the specific chondroitin sulfate core protein gene under study is that of chicken embryo sternal cartilage. In the case of laminin, the objectives are to characterize the molecular anatomy of that glycoprotein, describing the relationships of the polypeptide subunits in the entire molecule as well as the relationships of the oligosaccharides to the polypeptides. The subunits will be isolated and their compositions and structures determined, including the location and structures of the major carbohydrate substituents. The interactions between the subunits in the molecule will be examined by selective dissociation of disulfide linkages and by immuno-electron microscopy, using specific antibodies to each subunit. In the case of the gene which encodes for core protein, the cDNA structure will be determined, and genomic clones will be isolated from a library of chicken DNA, followed by determination of the sequence of the genomic DNA. Parallel studies of core protein mRNA expression in a well-characterized differentiating system, the chick embryo limb bud, will be done as well as determination of the copy number of the gene in chicken tissues. A human genomic library will also be screened with either the chicken cDNA probe or genomic chicken DNA. These two projects encompass the scientific disciplines of biochemistry and molecular biology, utilizing contemporary methods to discern protein structure, carbohydrate structure, and gene structure. The health-relatedness of the research applies to the general area of acquired and hereditary connective tissue diseases.

这是一个竞争性的继续要求， 支持. 拟议研究的主要重点是结构， 胞外大分子的功能和生物学表达 基质，检查层粘连蛋白的蛋白质和碳水化合物结构， 硫酸软骨素蛋白聚糖核心蛋白的基因结构。 层粘连蛋白是从小鼠EHS肿瘤中分离出来的， 所研究的硫酸盐核心蛋白基因是鸡胚胸骨硫酸盐核心蛋白基因 软骨 在层粘连蛋白的情况下，目标是表征分子结构。 糖蛋白的解剖，描述了 整个分子中的多肽亚基以及 将寡糖转化为多肽。 亚单位将被隔离 确定了它们的成分和结构， 和主要碳水化合物取代基的结构。 的相互作用 分子中亚基之间的关系将通过选择性的 二硫键解离和免疫电子显微镜，使用 每个亚单位的特异性抗体。 在编码核心蛋白的基因的情况下， 将被确定，基因组克隆将从一个文库中分离， 鸡DNA，然后测定基因组DNA的序列。 平行研究核心蛋白mRNA表达在一个良好表征的 分化系统，鸡胚肢芽，将以及 确定鸡组织中基因的拷贝数。 人类 基因组文库也将用鸡cDNA探针或 鸡的基因组DNA 这两个项目包括生物化学和生物化学的科学学科， 分子生物学，利用现代方法识别蛋白质 结构、碳水化合物结构和基因结构。 的 研究的健康相关性适用于获得性疾病的一般领域 和遗传性结缔组织疾病。