Characterization of Plasma Membrane Vitamin D Receptor

质膜维生素 D 受体的表征

• 批准号: 6460505
• 负责人: Susan Ellen Safford
• 金额: $ 22.95万
• 依托单位: LINCOLN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-17 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6460505
• 关键词: 1,25 dihydroxycholecalciferol; calcium flux; calcium indicator; cell membrane; confocal scanning microscopy; enzyme activity; functional /structural genomics; gene expression; immunocytochemistry; intermolecular interaction; minority institution research support; molecular cloning; protein kinase C; protein structure function; radiotracer; scintillation counter; tissue /cell culture; vitamin D receptors

DESCRIPTION (provided by applicant): Historically Black Colleges and Universities like Lincoln University are a major source of under represented students graduating with science degrees . Successful candidates to graduate and professional schools often have research experience obtained during their undergraduate years. To increase the competitiveness of our undergraduate students it is desirable to develop a competitive research program at Lincoln University. Therefore, the PI is seeking the research experience described below. This research will provide Lincoln students and PI with projects involving physiology, endocrinology, and molecular biology. The research plan focuses on characterization of a candidate mammalian plasma membrane vitamin D receptor (pmVDR). The best described active vitamin D metabolite, 1,25(OH)2D3, operates through nuclear receptor-mediated and plasma membrane- initiated mechanisms that are pharmacologically separable. The identity of a nuclear receptor is well documented, but the identity of an unequivocal membrane receptor for 1,25(OH)2D3 remains unknown. A 66kD protein from chicken intestine basal lateral membrane has been isolated by Nemere and colleagues and identified as a candidate receptor. The PI and collaborator have identified a candidate binding protein for the pmVDR whose N terminus is identical to that of the 66kD protein. They hypothesize that this recently identified l pmVDR participates in the generation of rapid responses in target cells to 1,25(OH)2D3. This application describes studies that will permit the evaluation of various expression constructs and transfected cell lines for the development of an optimal system for studying the physiology of the candidate pmVDR. Specifically, they will measure differences in the levels of rapid responses to 1,25(OH)2D3 in transfected cells compared to nontransfected controls. They will study tissue expression of the candidate pmVDR using various antibodies raised against different lengths of the sequence and use bioinformatics to help determine some of the proteins potential functions. These results might help lead to the development of new bioactive sterols with improved therapeutic potential and fewer side effects such as hypercalcemia.

描述(由申请人提供)：历史上的黑人学院和 像林肯大学这样的大学是代表不足的主要来源 获得理科学位毕业的学生。成功的应届毕业生 专业学校通常在他们的研究过程中获得研究经验 读本科的几年。提高本科生的竞争力 学生们希望在林肯大学制定一个有竞争力的研究项目 大学。因此，PI正在寻求所描述的研究经验 下面。这项研究将为林肯学生和PI提供项目 涉及生理学、内分泌学和分子生物学。研究计划 重点研究了一种哺乳动物质膜维生素D的候选基因 受体(PmVDR)。描述最好的活性维生素D代谢物1，25(OH)2D3， 通过核受体介导和质膜启动 这些机制在药物上是可以分离的。核子的身份 受体被很好地记录下来，但明确的膜的身份 1，25(OH)2D3的受体尚不清楚。鸡肠道中的一种66kD蛋白 Nemere和他的同事已经分离出基底膜和侧膜 被确定为候选受体。PI和合作者已经确定了一个 N端与pmVDR相同的候选结合蛋白 66kD蛋白。他们推测，这是最近发现的L pmVDR 在靶细胞中参与产生快速反应以 1，25(OH)2D3.本申请描述了将允许评估的研究 各种表达载体和转基因细胞系的研究进展 用于研究候选PMVDR的生理学的最佳系统。 具体地说，他们将衡量快速反应水平的差异 1，25(OH)2D3转染组与未转染组比较。他们会 用不同抗体研究候选pmVDR的组织表达 比对不同长度的序列并使用生物信息学来帮助 确定一些蛋白质的潜在功能。这些结果可能会有所帮助 导致新的生物活性甾醇的开发和治疗的改进 潜在的和更少的副作用，如高钙血症。