MOLECULAR PATHOLOGY AND PATHOGENESIS OF X-ADRENOLEUKODYSTROPHY

X-肾上腺脑白质营养不良的分子病理学和发病机制

• 批准号: 6296763
• 负责人: PAUL A WATKINS
• 金额: $ 32.52万
• 依托单位: HUGO W. MOSER RES INST KENNEDY KRIEGER
• 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 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6296763
• 关键词: acyl coA; adrenoleukodystrophy; antibody; disease /disorder etiology; enzyme activity; fatty acid metabolism; gene targeting; genetic disorder; genetic mapping; human tissue; laboratory rabbit; ligase; long chain fatty acid; membrane transport proteins; molecular cloning; molecular pathology; nucleic acid sequence; peroxisome; phenotype; protein structure function; sex linked trait; site directed mutagenesis; tissue /cell culture; yeast two hybrid system

The long-term objective of this project is to understand the molecular pathology and pathogenesis of X-linked adrenoleukodystrophy (ALD), a fatal neurodegenerative disorder. Defective peroxisomal activation of very long-chain fatty acids (VLCFA) by very long-chain fatty acyl-CoA synthetase (VLCS) causes elevated VLCFA levels in ALD patients. The product of the gene shown to be defective in ALD (ALDP) is a peroxisomal membrane protein that is a member of the ATP-binding cassette (ABC) membrane Transporter protein family and does not structurally resemble Acyl-CoA synthetases. Although ALDP is clearly required for normal degradation of VLCFA, it does not have VLCS activity, and neither its biochemical function nor its role in ALD have been elucidated. However, mutations in this protein lead to devastating clinical pathology. To develop clinical therapies and to study the pathophysiology of this disease, both genetic and biochemical studies of ALDP function will be used. The VLCS gene will be cloned and its gene product characterized in order to elucidate the relationship of VLCS, previously thought to be the defect in ALD, to ALDP. Two types of mutagenesis strategies, random mutagenesis and disruption of specific regions of the regions of the protein, will be used to analyze the structure and function of ALDP. Because ABC proteins often require interaction with other proteins for function, genetic and biochemical approaches will be used to elucidate ALDP-protein interactions. Finally, the effect of ALDP on VLCFA metabolism in several cell types (including cells of neural origin) and in model membranes will examined. These studies will lead to a better understanding of ALDP function and the precise roles of ALDP and VLCFA in the pathogenesis of ALD and perhaps provide clues for the phenotypic variability characteristic of ALD.

该项目的长期目标是了解分子 X连锁肾上腺脑白质营养不良（ALD）的病理学和发病机制 致命的神经退行性疾病。 过氧化物酶体激活缺陷 由极长链脂肪酰基辅酶A产生的极长链脂肪酸（VLCFA） 合成酶 (VLCS) 会导致 ALD 患者 VLCFA 水平升高。 这 ALD (ALDP) 缺陷基因的产物是过氧化物酶体 膜蛋白是 ATP 结合盒 (ABC) 的成员 膜转运蛋白家族，结构不相似 酰基辅酶A合成酶。 尽管 ALDP 显然是正常所必需的 VLCFA 的降解，它不具有 VLCS 活性，也不具有 生化功能及其在 ALD 中的作用尚未阐明。 然而， 这种蛋白质的突变会导致毁灭性的临床病理。 到 开发临床疗法并研究其病理生理学 疾病，ALDP 功能的遗传和生化研究将是 用过的。 VLCS基因将被克隆并鉴定其基因产物 为了阐明 VLCS 的关系，以前认为 ALD 的缺陷，变为 ALDP。 两种类型的诱变策略，随机 特定区域的诱变和破坏 蛋白质，将用于分析 ALDP 的结构和功能。 因为 ABC 蛋白通常需要与其他蛋白相互作用 将使用功能、遗传和生化方法来阐明 ALDP-蛋白质相互作用。 最后是ALDP对VLCFA的影响 多种细胞类型（包括神经源细胞）的新陈代谢 将在模型膜中进行检查。 这些研究将带来更好的结果 了解 ALDP 功能以及 ALDP 和 VLCFA 的确切作用 ALD 的发病机制，并可能为表型提供线索 ALD 的变异特征。