MOLECULAR STUDIES ON INTESTINAL MUCIN AND ITS FUNCTION

肠粘蛋白及其功能的分子研究

• 批准号: 2134006
• 负责人: DAVID R MACK
• 金额: $ 8.83万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2134006
• 关键词: Escherichia coli; animal genetic material tag; antibody; carbohydrate structure; gastrointestinal system; genetic library; guinea pigs; host organism interaction; human genetic material tag; molecular cloning; molecular genetics; mucins; nucleic acid sequence; oligosaccharides; protein structure function; tissue /cell culture

Mucins are important molecules whose strategic position between the intestinal luminal contents and mucosal epithelial cells has suggested a number of important biological roles including the protection against adhesion by enteropathogens. Adhesion of bacteria to epithelial cells is crucial for subsequent toxin delivery, colonization and invasion. A number of studies have shown that mucin inhibits bacterial adhesion and evidence has suggested that the capacity of mucin to inhibit bacterial adhesion is altered in animal models of human diseases. Further analysis of the mucin molecule will be restrained by the limitations of proteins (apomucins) has been available through the application of cDNA cloning strategies which represent a significant advance in the field. Whereas longer sequences of the MUC2 intestinal mucin gene have been forthcoming the same is not true of the MUC3 intestinal mucin gene. A cell line has been induced to overexpress MUC3 mRNA and generation of a cDNA library will allow for cloning and sequencing the MUC3 mucin gene. With the alteration of the cell line to overexpress either MUC2 or MUC3 the role of distinct mucins in bacterial adhesion will be evaluated. This will be accomplished through both the comparison of direct adhesion to the cell subcultures and by purifying the different mucins for evaluation of mucins and their constituents in the process of inhibition of bacterial adhesion. The determination of bacterial adhesion to mucin constituents offers new and important strategies for which to facilitate bacterial clearance from the intestinal tract. Finally, animal homologues to intestinal mucin genes will be identified. The knowledge of these sequences in animals may have important consequences to our understanding of the biological significance of the various domains contained within human mucins. The studies will be performed at a major medical center under the research direction of a world leader in the molecular aspects of mucins. The candidate's proposed studies would be extension of studies completed to date using molecular techniques that will propel this research well into the future.

粘蛋白是重要的分子，其战略地位在 肠腔内容物和粘膜上皮细胞表明 许多重要的生物学作用，包括防止 肠道病原体的粘附。 细菌与上皮细胞的粘附 对于随后的毒素输送、定植和入侵至关重要。 一个 大量研究表明，粘蛋白可以抑制细菌粘附， 有证据表明粘蛋白具有抑制细菌的能力 在人类疾病的动物模型中，粘附力发生了改变。 进一步分析 粘蛋白分子的功能会受到蛋白质的限制 (apomucins) 已通过 cDNA 克隆获得 代表该领域重大进步的策略。 然而 MUC2肠粘蛋白基因的更长序列即将问世 MUC3肠粘蛋白基因的情况则不然。 一个细胞系有 诱导过度表达 MUC3 mRNA 并生成 cDNA 文库 将允许克隆和测序 MUC3 粘蛋白基因。 随着 改变细胞系以过表达 MUC2 或 MUC3 的作用 将评估细菌粘附中的不同粘蛋白。 这将 可以通过直接粘附力的比较来完成 细胞传代培养物并通过纯化不同的粘蛋白来评估 粘蛋白及其成分在抑制细菌过程中的作用 附着力。 细菌对粘蛋白成分粘附的测定 提供了促进细菌生长的新的重要策略 从肠道清除。 最后，动物同源物 将鉴定肠道粘蛋白基因。 这些知识 动物中的序列可能对我们的理解产生重要影响 内包含的各个域的生物学意义 人类粘蛋白。 研究将在主要医疗中心进行 在分子方面世界领先者的研究方向下 粘蛋白。 候选人提议的研究将是 迄今为止使用分子技术完成的研究将推动 这项研究很适合未来。