FATTY ACID TRANSPORT--STRUCTURAL AND CELL BIOLOGY

脂肪酸运输——结构和细胞生物学

• 批准号: 6847167
• 负责人: JAMES Athur HAMILTON
• 金额: $ 22.65万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-26 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6847167
• 关键词: adipocytes; albumins; blood lipoprotein; calcium flux; calorimetry; fatty acid binding protein; fatty acid metabolism; fatty acid transport; fluorescence spectrometry; intermolecular interaction; lipid bilayer membrane; long chain fatty acid; membrane activity; membrane lipids; membrane model; membrane permeability; nuclear magnetic resonance spectroscopy; phospholipids; protein structure function; saturated fatty acids; stop flow technique; structural biology; tissue /cell culture; video microscopy

Fatty acids are essential nutrients of human cells but are also central players, if not causative agents, in several common diseases, including myocardial ischemia, diabetes and obesity. Acutely elevated leels of FA following an ischemic event disturb membrane-related functions such as contraction, excitability, rhythm and cell viability. Chronically elevated acids may be a direct causative agent in type II diabetes. Less common genetic disorders of fatty acid oxidation such as adrenoleukodystrophy (ALD) are characterized by accumulation of specific fatty acids, very long chain saturated fatty acids (VLCFA). This project uses state-of-the- art methods in both structural and cell biology to describe structural and dynamic aspects of fatty acid binding interactions and transport in plasma, cell membranes, and serum albumin by differential scanning calorimetry and 13C NMR and fluorescence spectroscopy, and studies the metabolic fate of VLCFA in cells of ALD patients by novel 13C NMR approaches. The second aim determines the complete tertiary structure in solution of intracellular fatty acid and lipid binding proteins (FABP); these structures provide a basis for elucidating molecular details of protein dynamics and ligand binding. Binding of less abundant ligands such as immediate products of fatty acid metabolism will be studied to elucidate new potential functions of FABP. The third aim studies the transport of fatty acids in membranes and tests both major hypotheses of fatty acid transport through the lipid bilayer, passive diffusion and protein-mediated transporter. This aim will use a combination of fluorescence probes to quantitative the adsorption and transmembrane movement steps of fatty acid transport in cells. Our studies of the fundamental and complex physical interactions and properties of fatty acids will prove indispensable knowledge for interpreting the behavior of fatty acids in physiological systems and delineating their precise roles in human diseases.

脂肪酸是人体细胞的必需营养素，但也是几种常见疾病（包括心肌缺血、糖尿病和肥胖症）的核心参与者（即使不是致病因素）。缺血事件后 FA 水平急剧升高会扰乱膜相关功能，例如收缩、兴奋性、节律和细胞活力。长期升高的酸可能是 II 型糖尿病的直接致病因素。不太常见的脂肪酸氧化遗传性疾病，例如肾上腺脑白质营养不良 (ALD)，其特征是特定脂肪酸、超长链饱和脂肪酸 (VLCFA) 的积累。该项目使用结构和细胞生物学领域最先进的方法，通过差示扫描量热法、13C NMR 和荧光光谱来描述脂肪酸结合相互作用以及血浆、细胞膜和血清白蛋白中转运的结构和动态方面，并通过新型 13C NMR 方法研究 ALD 患者细胞中 VLCFA 的代谢命运。第二个目标是确定细胞内脂肪酸和脂质结合蛋白（FABP）溶液中的完整三级结构；这些结构为阐明蛋白质动力学和配体结合的分子细节提供了基础。将研究不太丰富的配体（例如脂肪酸代谢的直接产物）的结合，以阐明 FABP 的新潜在功能。第三个目标是研究脂肪酸在膜中的转运，并测试脂肪酸通过脂质双层、被动扩散和蛋白质介导的转运蛋白转运的两个主要假设。该目标将使用荧光探针组合来定量细胞中脂肪酸转运的吸附和跨膜运动步骤。我们对脂肪酸的基本和复杂的物理相互作用和特性的研究将证明，这对于解释脂肪酸在生理系统中的行为和描述其在人类疾病中的精确作用是不可或缺的知识。