Biomolecular Analysis of Proteins in Visual Disease

视觉疾病中蛋白质的生物分子分析

• 批准号: 7195581
• 负责人: ESTHER Elissa BISWAS-FISS
• 金额: $ 23.25万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7195581
• 关键词: 220kDa rod outer segment rim protein; ATP phosphohydrolase; ATP-Binding Cassette Transporters; Affect; Age related macular degeneration; Area; Binding; Biochemical; Blindness; Circular Dichroism; Clinical; Complementary DNA; Complex; Coupling; Defect; Development; Disease; Environment; Extracellular Domain; Fluorescence; Fluorescence Anisotropy; Funding; Future; Gene Mutation; Genes; Genetic; Genotype; Grant; Hydrolysis; Individual; Inherited; Knowledge; Label; Laboratories; Lead; Length; Ligands; Lipofuscin; Localized; Macular degeneration; Membrane; Molecular; Molecular Analysis; Molecular Conformation; Mutation; Nucleotides; Phenotype; Photoreceptors; Property; Protein Analysis; Proteins; Public Health; Recombinants; Retina; Retinal; Retinal Degeneration; Retinitis Pigmentosa; Retinoids; Ribonucleotides; Role; SHFM1 gene; Secretory Vesicles; Severities; Site-Directed Mutagenesis; Specificity; Spectrum Analysis; Stargardt' s disease; Structure; Substrate Interaction; Systems Analysis; Testing; Thinking; United States; Vertebrate Photoreceptors; Vesicle; Visual; Visual impairment; Yeasts; aging population; base; clinical phenotype; disease phenotype; extracellular; mutant; outcome forecast; photoreceptor disc; polypeptide; predictive modeling; protein function; response

DESCRIPTION (provided by applicant): The ABCA4 gene encodes a retina specific ATP binding cassette (ABC) transporter, ABCR, which is localized to the rod and cone photoreceptor outer segments. ABCR is thought to function in the transport of retinal derivatives across photoreceptor disk membranes. Mutations in the ABCA4 gene lead to several inherited forms of retinal degeneration (RD); including Stargardt disease, cone-rod dystrophy (arCRD), fundus flavimaculatus (FFM) and age related macular degeneration (ARMD). These degenerations represent a broad spectrum of clinical phenotypes, all associated with accumulations of lipofuscin, which is thought to be in part due to defects in transport by ABCR. Our overall aim is to investigate the mechanism of action of ABCR protein and how genetic mutations observed in Stargardt disease and other RDs influence ABCR structure and function so that new and more precise therapies could be developed in the near future. Our proposed studies will focus on two areas, first the function of specific domains of ABCR and second the function of the protein as a whole. We plan to test the hypothesis that RD associated mutations, in the extracellular (ECD) or nucleotide binding (NBD) domains lead to conformational changes, which in turn influence protein-protein or protein-ligand interactions. This will help us to develop a predictive model for the likely effect of a given mutation on structural and functional changes in these domains. Second, we shall test the hypothesis that retinal or retinal derivatives interact directly with the domains of ABCR in a membrane bound environment. We shall express full-length ABCR directly in membrane vesicles in order to test this hypothesis. These studies will provide us an avenue to investigate the effects of RD associated mutations on the ABCR molecule as a whole. Macular degenerations are the leading cause of blindness in the United States, and visual impairment is an increasing public health issue, particularly with our aging population. Several types of degeneration, affecting both the young and the old, have been associated with mutations in the ABCR gene. These studies will lead to a more accurate understanding of the involvement ABCR in the development of macular degenerations, such as Cone Rod Dystrophy, Stargardt disease, and Age Related Macular Degeneration. This will enable us to develop a more accurate prognosis of those individuals identified with mutations in the ABCR gene, and ultimately, the development of better therapies. Overall, our studies seek to bridge the knowledge obtained through clinical genetics, and the consequences of these mutations at the molecular level -- thereby enabling us to develop a clinical genotype-biochemical phenotype correlation.

描述(申请人提供)：ABCA4基因编码视网膜特异性三磷酸腺苷结合盒(ABC)转运体ABCR，定位于视杆和视锥感光细胞外段。ABCR被认为在视网膜衍生物通过光感受器盘膜的运输中起作用。ABCA4基因突变导致多种遗传性视网膜变性(RD)，包括Stargardt病、视锥视杆细胞营养不良(ArCRD)、黄斑眼底病(FFM)和老年性黄斑变性(ARMD)。这些变性代表了广泛的临床表型，都与脂褐素的积聚有关，这被认为部分是由于ABCR运输的缺陷。我们的总体目标是研究ABCR蛋白的作用机制，以及在Stargardt病和其他RDS中观察到的基因突变如何影响ABCR的结构和功能，以便在不久的将来开发新的更精确的治疗方法。我们建议的研究将集中在两个领域，第一，ABCR特定结构域的功能；第二，蛋白质作为一个整体的功能。我们计划测试这一假设，即细胞外(ECD)或核苷酸结合(NBD)区域的RD相关突变会导致构象变化，进而影响蛋白质-蛋白质或蛋白质-配体的相互作用。这将帮助我们开发一个预测模型，预测给定突变对这些结构域的结构和功能变化的可能影响。其次，我们将检验这一假设，即视网膜或视网膜衍生物在膜结合的环境中直接与ABCR结构域相互作用。为了验证这一假说，我们将在膜泡中直接表达全长ABCR。这些研究将为我们提供一个途径来研究RD相关突变对整个ABCR分子的影响。黄斑变性是美国致盲的主要原因，视力障碍是一个日益严重的公共健康问题，特别是在我们人口老龄化的情况下。影响年轻人和老年人的几种类型的退行性疾病与ABCR基因的突变有关。这些研究将有助于更准确地理解ABCR在黄斑变性的发生发展中的作用，如锥状杆营养不良、Stargardt病和年龄相关性黄斑变性。这将使我们能够对那些被确认为ABCR基因突变的个体进行更准确的预后，并最终开发出更好的治疗方法。总体而言，我们的研究试图在通过临床遗传学获得的知识和这些突变在分子水平上的后果之间架起桥梁--从而使我们能够发展临床基因型与生化表型的相关性。