Characterization of Plasma Membrane Vitamin D Receptor

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性不足的主要来源 获得科学学位的学生。成功毕业的候选人 专业学校通常有在其学习期间获得的研究经验 本科年。提高本科生竞争力 学生们希望在林肯大学开发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 专注于候选哺乳动物质膜维生素 D 的表征 受体（pmVDR）。最有描述的活性维生素 D 代谢物 1,25(OH)2D3， 通过核受体介导和质膜启动起作用 药理学上可分离的机制。核的身份 受体已被充分记录，但膜的身份明确 1,25(OH)2D3 的受体仍然未知。来自鸡肠的 66kD 蛋白质 Nemere 及其同事分离出了基底侧膜 被鉴定为候选受体。 PI 和合作者已经确定了 pmVDR 的候选结合蛋白，其 N 末端与 66kD 蛋白质。他们推测最近发现的 l pmVDR 参与靶细胞快速反应的产生 1,25(OH)2D3。该应用程序描述了允许评估的研究 用于开发的各种表达构建体和转染细胞系 研究候选 pmVDR 生理学的最佳系统。 具体来说，他们将衡量快速反应水平的差异 与未转染的对照相比，转染细胞中的 1,25(OH)2D3。他们会 使用产生的各种抗体研究候选 pmVDR 的组织表达 针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，并改善治疗效果 潜力和较少的副作用，如高钙血症。

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.