INTESTINAL VITAMIN A-BINDING PROTEINS

肠道维生素 A 结合蛋白

• 批准号: 2141208
• 负责人: ELLEN LI
• 金额: $ 14.7万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-15 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2141208
• 关键词: Escherichia coli; affinity labeling; chemical binding; chemical models; computer simulation; fluorescence spectrometry; gastrointestinal nutrient absorption; laboratory rat; membrane model; nuclear magnetic resonance spectroscopy; nutrition related tag; protein structure; recombinant proteins; retinoid binding proteins; retinoids; site directed mutagenesis; vitamin metabolism

Vitamin A is an essential nutrient required for growth, reproduction, differentiation of epithelial tissues and vision. The mechanism by which dietary retinoids are absorbed and converted to various metabolites is complex. Specific retinoid binding proteins have been implicated in mediating the absorption, intracellular trafficking and the action of vitamin A. In particular, we are interested in the role of cellular retinol binding protein II (CRBP II), which is present exclusively in the small intestine in the adult animal, suggesting that it is uniquely adapted to the intestinal absorption and metabolism of retinol. CRBP II is closely homologous to cellular retinol binding protein (CRBP), which in contrast to CRBP II, is found in many epithelial tissues. Monospecific ligands that bind to only one protein, would be extremely useful for dissecting the metabolic roles of the retinoid binding proteins, and could potentially lead to the design of new drugs. To identify or design such ligands requires further understanding of the molecular details of binding interactions for these proteins. We propose to study these interactions as they occur in solution by using recombinant proteins expressed in E. coli, fluorescence and nuclear magnetic resonance spectroscopy, photoaffinity labeling, computer-assisted molecular modeling and mutagenesis experiments. We also propose to apply techniques developed to study the intracellular retinol binding proteins towards the analysis of intracellular retinoic acid binding proteins, including cellular retinoic acid binding protein (CRABP), and the nuclear retinoic acid binding proteins (including members of the RAR and RXR family).

维生素A是生长、繁殖、 上皮组织和视觉的分化。 其机制 饮食中的类维生素A被吸收并转化为各种代谢物 复杂的。 特定的类视黄醇结合蛋白与 介导吸收、细胞内运输和作用 维生素 A。我们特别对细胞的作用感兴趣 视黄醇结合蛋白 II (CRBP II)，仅存在于 成年动物的小肠，表明它是独特的 适应肠道对视黄醇的吸收和代谢。 CRBP II 与细胞视黄醇结合蛋白 (CRBP) 密切同源， 与 CRBP II 不同，CRBP II 存在于许多上皮组织中。 仅与一种蛋白质结合的单特异性配体将是极其重要的 对于剖析类维生素A结合的代谢作用很有用 蛋白质，并可能导致新药物的设计。 到 识别或设计此类配体需要进一步了解 这些蛋白质结合相互作用的分子细节。 我们建议 通过使用来研究溶液中发生的这些相互作用 在大肠杆菌中表达的重组蛋白、荧光和核 磁共振波谱、光亲和标记、 计算机辅助分子建模和诱变实验。 我们 还建议应用开发的技术来研究细胞内 视黄醇结合蛋白用于细胞内视黄酸的分析 酸结合蛋白，包括细胞视黄酸结合蛋白 （CRABP）和核视黄酸结合蛋白（包括 RAR 和 RXR 家族的成员）。