Biochemical dissection of amyloid precursor protein transport from the Golgi

高尔基体淀粉样前体蛋白转运的生化剖析

• 批准号: 8904033
• 负责人: Daniel W. Sirkis
• 金额: $ 5.42万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8904033
• 关键词: Adaptor Signaling Protein; Address; Affect; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein Precursor; Arl proteins; Biochemical; Biological Assay; Biological Process; California; Cell surface; Cells; Cellular biology; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Cytoplasmic Protein; Cytosol; Disease; Dissection; Endoplasmic Reticulum; Endosomes; Event; Family; GTP-Binding Proteins; Generations; Genes; Genetic study; Goals; Golgi Apparatus; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Healthcare Systems; Human Genetics; Hydrolysis; Incidence; Integral Membrane Protein; Late Onset Alzheimer Disease; Learning; Light; Location; Mediating; Membrane Proteins; Mentors; Metabolism; Mission; Molecular Biology; Monitor; Mutagenesis; National Institute on Aging; Nerve Degeneration; Neurodegenerative Disorders; Outcome; Pathogenesis; Pathway interactions; Phospholipase; Postdoctoral Individual National Research Service Award; Process; Production; Protein Export Pathway; Proteins; RNA Interference; Research; Research Project Grants; Risk Factors; Role; Site; Sorting - Cell Movement; Techniques; Testing; Therapeutic Intervention; Time; Training; United States; Universities; Variant; Vesicle; Work; career; experience; genetic risk factor; genome editing; novel; protein metabolism; protein transport; public health relevance; receptor; reconstitution; residence; secretase; sortilin; trafficking; trans-Golgi Network

 DESCRIPTION (provided by applicant): This is a proposal for an F32 award for Dr. Daniel Sirkis, a postdoctoral associate in the Department of Molecular and Cell Biology at the University of California, Berkeley. Dr. Sirkis is a cell biologist focusing on the intracellular trafficking of proteins implicated in neurodegenerative disease. This F32 will provide Dr. Sirkis with the support required to accomplish the following goals: (i) to gain experience in the biochemical reconstitution of vesicle trafficking events; (ii) to learn state-of-the-art genome-editing techniques, such as CRISPR/Cas9-mediated mutagenesis; (iii) to use these techniques to clarify the relationship between the intracellular sorting of the amyloid precursor protein (APP and the generation of its proteolytic cleavage product, amyloid-ß (Aß), both of which have been implicated in Alzheimer's disease (AD); and (iv) to gain additional experience in scientific communication and mentoring that will be required for a smooth transition to an independent research career. Dr. Sirkis has chosen to carry out his postdoctoral studies under the sponsorship of Dr. Randy Schekman, a cell biologist with deep expertise in vesicle trafficking and biochemical reconstitution. The proposed research project focuses on the intracellular sorting of APP at the trans-Golgi network (TGN). Because the TGN has been implicated as a major site of Aß generation, this work has the potential to shed light on pathogenic processes involved in AD. In Aim 1, Dr. Sirkis will determine whether small GTP- binding proteins found to influence Aß production are involved in APP trafficking at the TGN. In Aim 2, Dr. Sirkis will determine whether the transmembrane protein SorLA, a known risk factor for late-onset AD, influences Aß production by facilitating APP transport from either the TGN or endosomes. In Aim 3, Dr. Sirkis will determine whether novel AD risk factor phospholipase D3 influences Aß production via the endoplasmic reticulum or the TGN. The overarching goals of this project are (i) to characterize how novel, Golgi-localized, GTP-binding proteins regulate the transport of APP at the TGN; and (ii) clarify how two genetic risk factors for late-onset AD, both of which are membrane proteins, affect the trafficking of APP. If successful, the proposed studies are expected to increase our understanding of APP sorting and Aß generation, and by extension, elucidate biological processes underpinning AD. Importantly, Dr. Sirkis' F32 training will prepare him for an independent research career that focuses on the intracellular trafficking of proteins involved in neurodegeneration.

 描述（由适用提供）：这是丹尼尔·西尔基斯（Daniel Sirkis）博士的F32奖的提案，丹尼尔·西尔基斯（Daniel Sirkis）博士是加利福尼亚大学伯克利分子和细胞生物学系的博士后学员。 Sirkis博士是一名细胞生物学家，专注于神经退行性疾病中实施的蛋白质的细胞内运输。该F32将为Sirkis博士提供实现以下目标所需的支持：（i）获得囊泡贩运事件的生化重构经验； （ii）学习最先进的基因组编辑技术，例如CRISPR/CAS9介导的诱变； （iii）使用这些技术来阐明淀粉样蛋白的细胞内分类（APP及其蛋白水解裂解产物的产生，淀粉样酶（Aß），这两种都暗示了在阿尔茨海默氏病（AD）和（IV）中，在科学方面的经验和精神上的经验都可以使他们能够平稳地进行跨越的经验。被选为兰迪·施克曼（Randy Schekman）的赞助，这是一名在囊泡贩运和生物化学重构方面的专业知识AIM 1，Sirkis博士将确定发现会影响Aß产生的小型GTP结合蛋白是否参与TGN的应用程序运输。在AIM 2中，Sirkis博士将确定跨膜蛋白Sorla是迟到AD的已知危险因素是否通过支持来自TGN或内体的应用传输来影响Aß产生。在AIM 3中，Sirkis博士将确定新颖的AD风险因子磷脂酶D3是通过内质网还是TGN影响Aß产生的。该项目的总体目标是（i）来表征新颖，golgi定位的GTP结合蛋白如何调节APP在TGN的运输； （ii）阐明迟到AD的两个遗传危险因素如何影响APP的运输。成功的研究预计将增加我们对应用程序分类和Aß产生的理解，并扩大阐明AD的生物学过程。重要的是，Sirkis博士的F32培训将为他准备独立研究职业，重点是涉及神经退行性的蛋白质的细胞内贩运。