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: 6395930
• 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: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6395930
• 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连锁肾上腺素的病理学和发病机理，A 致命的神经退行性疾病。 过氧化物酶体激活 非常长链脂肪酸（VLCFA）非常长链脂肪酰基辅酶A 合成酶（VLC）导致ALD患者的VLCFA水平升高。 这 该基因在ALD中有缺陷（ALDP）的产物是过氧化物酶体 膜蛋白是ATP结合盒（ABC）的成员 膜转运蛋白蛋白家族，与结构上不相似 酰基-COA合成酶。 虽然正常需要ALDP VLCFA的退化，它没有VLCS活动，也没有 阐明了生化功能或其在ALD中的作用。 然而， 该蛋白质的突变导致毁灭性的临床病理。 到 开发临床疗法并研究此的病理生理学 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，其基因产物具有特征 为了阐明VLC的关系，以前认为是 ALD中的缺陷到ALDP。 两种类型的诱变策略，随机 诱变和破坏该区域的特定区域 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质相互作用 功能，遗传和生化方法将用于阐明 ALDP蛋白质相互作用。 最后，ALDP对VLCFA的影响 几种细胞类型的代谢（包括神经来源的细胞）和 在模型膜中将检查。 这些研究将导致更好 了解ALDP功能以及ALDP和VLCFA的精确作用 在ALD的发病机理中，也许为表型提供了线索 ALD的可变性特征。