Characterization of Plasma Membrane Vitamin D Receptor

质膜维生素 D 受体的表征

• 批准号: 7066031
• 负责人: Susan Ellen Safford
• 金额: $ 23.66万
• 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 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7066031
• 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）：历史上的黑人大学和 像林肯大学这样的大学是代表不足的主要来源 学生获得科学学位的毕业。成功毕业的候选人 专业学校经常有研究经验 本科生。提高本科生的竞争力 学生希望在林肯制定竞争性研究计划 大学。因此，PI正在寻求描述的研究经验 以下。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 专注于候选哺乳动物质膜维生素D的表征 受体（PMVDR）。最好描述的活性维生素D代谢物，1,25（OH）2d3， 通过核受体介导的质膜和质膜进行操作 在药理学上可分开的机制。核的身份 受体有充分的文献记载，但明确的膜的身份 1,25（OH）2d3的受体仍然未知。鸡肉肠的66kD蛋白 Nemere及其同事已经分离了基底外侧膜 被识别为候选受体。 PI和合作者已经确定了 n末端与之相同的PMVDR的候选蛋白 66KD蛋白。他们假设这最近确定了L PMVDR 参与目标细胞中快速反应的产生 1,25（OH）2d3。该应用程序描述了将允许评估的研究 各种表达构建体和转染的细胞系 研究候选PMVDR的生理学的最佳系统。 具体来说，他们将衡量对快速响应水平的差异 与未转染的对照相比，转染的细胞中的1,25（OH）2d3。他们会的 研究候选PMVDR的组织表达使用各种抗体 在序列的不同长度上，并使用生物信息学来帮助 确定一些蛋白质潜在功能。这些结果可能有帮助 通过改进的治疗性发展新的生物活性固醇 潜力和较少的副作用，例如高钙血症。