LOCALIZATION AND PROTEIN INTERACTIONS OF FRATAXIN

Frataxin 的定位和蛋白质相互作用

• 批准号: 6393140
• 负责人: GEORGE R WILMOT
• 金额: $ 12.78万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6393140
• 关键词: Friedreich's ataxia; affinity chromatography; antibody formation; antibody specificity; antiserum; binding proteins; density gradient ultracentrifugation; human tissue; immunocytochemistry; immunoelectron microscopy; immunoprecipitation; intermolecular interaction; intracellular transport; laboratory mouse; laboratory rat; molecular pathology; neurogenetics; postmortem; protein purification; protein structure function; protein transport; structural genes; western blottings; yeast two hybrid system

The recent identification of the gene asssociated with Friedreich's ataxia (FA) has provided an exciting opportunity to learn more about the molecular pathenogenesis of this devastating, progressively debilitating disease. The gene, termed X25, was found to contain an expanded GAA trinucleotide repeat within the first intron in the vast majority of FA patients, which presumably leads ultimately to an absent or severely reduced level of its coded protein "frataxin". However, no direct studies have yet been reported on frataxin. Unffl this occurs, the cellular pathophysiology leading to the disease wffl remain a mystery, and the development of specific treatment strategies will be hindered. The experiments outined in this proposal are therefore directed toward understanding the normal cellular function of frataxin. In Specific Aim 1, we propose to develop and characterize affinity- purified polyclonal antisera which specifically recognize frataxin. Preliminary studies have already demonstrated that antibodies raised to recombinant frataxin recognize proteins in human tissue homogenates that may correspond to ftataxin. In Specific Aim 2, we will utilize the antibodies developed in Specific Aim 1 to determine the tissue, cellular, and subcellular distribution of frataxin in mice and human control tissues. Since protein function is largely determined by protein-prqtein interactions, we will attempt to identify frataxin- associated proteins in Specific Aim 3. In addition to these research activities, this proposal is being submitted as part of an overall career development plan. As such, additional clinical and didactic activities are outlined which will provide the principal investigator an excellent opportunity to continue his development as an academic neurologist and help to ensure his success as a research scientist.

最近与弗里德里希（Friedreich）相关的基因屁股的鉴定 共济失调（FA）提供了一个令人兴奋的机会来了解有关 这种毁灭性的分子疗法发生，逐渐 使人衰弱的疾病。该基因称为x25，被发现包含一个 扩展的GAA三核苷酸在广大内含子中重复 大多数FA患者，可能最终导致 其编码蛋白“ Frataxin”的缺乏或严重降低。 但是，尚无关于Frataxin的直接研究。 unffl 发生这种情况，导致疾病WFFL的细胞病理生理学 仍然是一个谜，并发展特定的治疗策略 将受到阻碍。 因此，该提案中提出的实验是指导的 了解frataxin的正常细胞功能。 在 特定目的1，我们建议发展和表征亲和力 - 纯化的多克隆抗血清，专门识别frataxin。 初步研究已经表明抗体提出了 重组Frataxin识别人体组织中的蛋白质 可能对应于ftataxin的匀浆。在特定的目标2中，我们 是否会利用特定目标1中开发的抗体来确定 小鼠和小鼠的组织，细胞和亚细胞分布 人类控制组织。 由于蛋白质功能在很大程度上取决于 蛋白质 - 普尔克特蛋白相互作用，我们将尝试确定frataxin- 特定目标3中的相关蛋白质。 除了这些研究活动外，还提交了该建议 作为整体职业发展计划的一部分。 因此，附加 概述了临床和教学活动，这将提供 首席调查员是继续他的 作为一名学术神经科医生的发展，并有助于确保他的成功 作为研究科学家。