MONOCLONAL ANTIBODIES AS PROBES TO VITAMIN D3 ACTION

单克隆抗体作为维生素 D3 作用的探针

• 批准号: 3158447
• 负责人: J WESLEY PIKE
• 金额: $ 16.12万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3158447
• 关键词: 1,25 dihydroxycholecalciferol; chickens; clone cells; enzyme mechanism; fibroblasts; human subject; laboratory mouse; laboratory rat; messenger RNA; monoclonal antibody; nucleic acid structure; protein metabolism; protein structure function; receptor

The diverse biologic actions of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) are thought to be mediated byintracellular receptor proteins which appear to operate at the level of the cell nucleus. We propose to continue our study of these unique polypeptides derived from both avian and mammalian sources utilizing immunologic as well as conventional probes. Investigation will focus upon four areas essential to our understanding of the molecular mechanism of 1,25(OH)2D3 action: 1) physical and biochemical properties of receptors, 2) intracellular biology of receptors and the role of 1,25(OH)2D3 3) functional characterization of receptors, and 4) molecular defects in receptors which confer cellular resistance to 1,25(OH)2D3. Physical and biochemical characterization will include evaluation of the two antigenically related forms of the avian receptor, topological mapping of both avian and mammalian receptors, determination of mammalian 1,25(OH)2D3-dependent receptor phosphoamino acid species, and isolation and sequence analysis of both avian and mammalian receptors. Aspects of the biology of 1,25(OH)2D3 receptors will include further evaluation of the intracellular origins of unoccupied forms, receptor synthesis and degradation, the effects of 1,25(OH)2D3 on cellualr levels of receptor and its mRNA, the nature and mechanism of receptor inactivation. Functional studies will include determination of potential receptor enzymatic activities, evaluation of teh functional significance of receptor modification, and analysis of receptor DNA sequence preference utilizing cloned vitamin D-dependent calbindin gene and receptor gene fragments. Evaluation of molecular abnormalities will involve the application of the above technology to the examination of abberrant human receptors from fibroblasts obtained from patients with reisitance to 1,25(OH)2D3 as well as from developed nonhuman variants. Collectively, the studies proposed here on normal receptors represent an integrated approach to the elucidation of the role of receptor in mediating various bilogical phenomena regulated by 1,25(OH)2D3. In addtion, they provide the basis for interpreting molecular defects in receptors which may be associated with vitamin D3-resistant disease states.

1，25-二羟维生素D3（1，25（OH）2D 3）的多种生物学作用是 被认为是由细胞内受体蛋白介导的， 在细胞核的水平上运作。 我们建议继续研究 这些独特的多肽来源于鸟类和哺乳动物来源 利用免疫学以及常规探针。 调查将 重点关注四个对我们理解分子生物学至关重要的领域。 1，25（OH）_2D_3的作用机理：1）1，25（OH）_2D_3的物理和生化性质 受体，2）受体的细胞内生物学和作用 3）受体的功能表征，和4）分子 受体缺陷，赋予细胞对1，25（OH）2D 3的抗性。 物理和生物化学表征将包括评价 鸟类受体的两种抗原相关形式，拓扑作图 鸟类和哺乳动物受体，确定哺乳动物 1，25（OH）2D 3依赖性受体磷酸氨基酸种类，以及分离和 鸟类和哺乳动物受体的序列分析。 方面 1，25（OH）2D 3受体的生物学研究将包括进一步评估 未占据形式的细胞内起源，受体合成和 降解，1，25（OH）_2D_3对细胞受体水平的影响， 它的mRNA，受体失活的性质和机制。 功能 研究将包括确定潜在的受体酶 活性，受体功能意义的评价 修饰和分析受体DNA序列偏好， 克隆的维生素D依赖性钙结合蛋白基因和受体基因片段。 分子异常的评估将涉及应用 上述技术用于检测来自 从对1，25（OH）2D 3具有抗性的患者中获得的成纤维细胞 从非人类进化的变异体中 总的来说，这些研究提出 在这里，正常受体代表了一个综合的方法， 阐明受体在介导各种生物学效应中的作用 由1，25（OH）2D 3调节的现象。 此外，它们还提供了 解释受体中的分子缺陷， 维生素D3抵抗疾病状态。