RETINOIDS, M6P/IGF II RECEPTOR & CELL GROWTH REGULATION

类维生素A、M6P/IGF II 受体

• 批准号: 2726421
• 负责人: JING X KANG
• 金额: $ 26.13万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2726421
• 关键词: SDS polyacrylamide gel electrophoresis; affinity chromatography; affinity labeling; animal tissue; autoradiography; binding sites; cell growth regulation; confocal scanning microscopy; electron microscopy; fluorescence microscopy; growth factor receptors; high performance liquid chromatography; immunoelectron microscopy; immunologic assay /test; immunoprecipitation; insulinlike growth factor; mannose 6 phosphate; polymerase chain reaction; protein purification; protein sequence; protein structure function; receptor binding; retinoid binding proteins; retinoids; western blottings

Both retinoids and the mannose 6-phosphate/insulin-like growth factor-II (M6P/IGF2) receptor have been shown to play fundamental roles in the control of cell growth in development and oncogenesis. Recent work has discovered that retinoic acid (RA) binds to the M6P/IGF2 receptor with high affinity, leading to alteration of the primary functions of the receptor and suppression of cell growth. These findings suggest that M6P/IGF2 receptor mediates a novel RA response pathway that may be important in cell growth regulation. However, the molecular mechanism underlying this pathway remains largely unknown. Thus, the overall goal of this proposal is to understand how RA interacts with the M6P/IGF2 receptor and the role of this interaction in cell growth regulation. The key questions that we wish to address are: 1) Where is the functional domain of the receptor protein responsible for retinoid binding? 2) What is the authentic biological consequence of the interaction? and 3) How do they work? Accordingly, the specific aims of this application are: (1) To identify the retinoid binding site of the M6P/IGF2 receptor; (2) To extend the understanding of the biological consequence of the RA-M6P/IGF2R interaction. (3) To identify the biochemical processes responsible for the biological effect of RA-M6P/IGF2R interaction. Our general approach is to create a mutant M6P/IGF2R that fails to bind RA and then assess differences of functional responses to RA between the cells expressing wild-type M6P/IGF2R and those with the mutant receptor. The principal methods to be used include protein mutagenesis, photoaffinity labeling, amino acid sequence analysis, ligand binding assay, immuno-detection, enzymatic assay and morphological analysis. Significance This project will provide fundamental information about how retinoids interact with the M6P/IGF2 receptor and shed light on the role of this novel retinoid-response pathway in regulation of cell growth. This knowledge may lead to novel or more effective therapeutic and preventive approaches for cancer and developmental defects.

类视黄素和甘露糖6-磷酸/胰岛素样生长 因子-II（M6P/IGF2）受体已显示出发挥基本的作用 控制细胞生长在发育和肿瘤发生中的作用。 最近的工作发现视黄酸（RA）与M6P/IGF2结合 具有高亲和力的受体，导致原发性改变 受体的功能和细胞生长的抑制。 这些 发现表明M6P/IGF2受体介导了新的RA响应 可能在细胞生长调节中很重要的途径。 但是， 该途径的分子机制在很大程度上仍然未知。 因此，该提议的总体目标是了解RA如何 与M6P/IGF2受体相互作用和这种相互作用的作用 在细胞生长调节中。 我们希望解决的关键问题 是：1）受体蛋白的功能结构域在哪里 负责类维生素性结合？ 2）什么是真实的生物学 互动的结果？ 3）它们如何工作？ 因此， 该应用程序的具体目的是：（1）识别类维生素 M6P/IGF2受体的结合位点； （2）扩展理解 RA-M6P/IGF2R相互作用的生物学后果。 （3）到 确定负责生物学的生化过程 RA-M6P/IGF2R相互作用的影响。 我们的一般方法是创建 一种突变的M6P/IGF2R，无法结合RA，然后评估 表达野生型细胞之间对RA的功能响应 M6P/IGF2R和具有突变体受体的人。 主要方法 使用包括蛋白质诱变，光性标记，氨基酸 序列分析，配体结合测定，免疫检测，酶促分析 测定和形态分析。 意义这个项目将提供有关的基本信息 类视黄素如何与M6P/IGF2受体相互作用并在 这种新型的类视视视感素反应途径在细胞调节中的作用 生长。 这些知识可能会导致新颖或更有效的治疗 以及癌症和发育缺陷的预防方法。