PORCINE SUBMAXILLARY MUCIN AND HUMAN NORRIN

猪颌下粘蛋白和人诺里蛋白

• 批准号: 6489969
• 负责人: ROBERT L HILL
• 金额: $ 31.93万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6489969
• 关键词: X ray crystallography; blindness; cell line; deafness; disulfide bond; expression cloning; genetic library; glycoproteins; laboratory rabbit; mental retardation; molecular assembly /self assembly; molecular site; mucins; nuclear magnetic resonance spectroscopy; posttranslational modifications; protein biosynthesis; protein folding; protein sequence; protein structure function; site directed mutagenesis; tissue /cell culture

The structure/function relationships of two related proteins will be examined. Porcine submaxillary mucin is a glycoprotein containing about 67 percent by weight carbohydrate. Mucins lubricate the respiratory, gastrointestinal and urogenital tracts and protect epithelial cells from dehydration and injury. Much of porcine submaxillary mucin structure has been elucidated, but not all, including how the monomeric mucin molecules assemble to form dimeric and multimeric mucins that are the mature, functional molecules. The half-cystines that form the inter- and intrachain disulfide bonds in dimers and multimers will be determined. Particular attention will be given to the structural motif CGLCG and its role in assembly of multimeric mucin. These studies will utilize site-directed mutagenesis and expression of mutant constructs in animal cells in culture. Similar studies will be performed with norrin, a 133 residue human protein that is defective in Norrie disease, an X-linked, recessive human neurological disorder characterized by blindness, deafness and mental retardation. This protein is very similar structurally to the domain in some mucins that forms disulfide- linked multimers. The half-cystines in norrin that form inter- and intrachain disulfide bonds will be determined by the same methods used with submaxillary mucin. Hopefully, these studies will reveal how the disulfide-rich domains in proteins in which they occur lead to oligomer formation.

两个相关蛋白质的结构/功能关系将是 考察 猪颌下粘蛋白是一种糖蛋白， 67%的碳水化合物 粘蛋白润滑呼吸道， 胃肠道和泌尿生殖道，并保护上皮细胞， 脱水和受伤。 大部分猪颌下粘蛋白结构 已经阐明，但不是全部，包括单体粘蛋白如何 分子组装形成二聚和多聚粘蛋白， 成熟的功能分子 半胱氨酸形成了 二聚体和多聚体中的链内二硫键将被 测定 将特别注意结构主题 CGLCG及其在多聚体粘蛋白组装中的作用。 这些研究将 利用定点诱变和突变构建体的表达 在培养的动物细胞中。 将进行类似的研究， norrin，一种在诺里氏病中有缺陷的133个残基的人类蛋白质， 一种X连锁隐性人类神经系统疾病，特征为 失明、失聪和智力迟钝。 这种蛋白质非常 在结构上类似于一些粘蛋白中形成二硫键的结构域， 连锁多聚体Norrin中的半胱氨酸， 链内二硫键将通过所用相同方法测定 颌下粘蛋白 希望这些研究能揭示 蛋白质中富含二硫键的结构域导致寡聚体 阵