Roles of PEN-2 and APH-1 in PS1 complex formation

PEN-2 和 APH-1 在 PS1 复合物形成中的作用

• 批准号: 6804546
• 负责人: WENJIE LUO
• 金额: $ 4.89万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-10-01 至 2005-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6804546
• 关键词: Alzheimer' s disease; Golgi apparatus; SDS polyacrylamide gel electrophoresis; amyloid proteins; cell line; endopeptidases; endoplasmic reticulum; enzyme activity; enzyme linked immunosorbent assay; enzyme mechanism; gene mutation; immunoprecipitation; intermolecular interaction; polymerase chain reaction; postdoctoral investigator; presenilin; protein folding; protein structure function; western blottings

DESCRIPTION (provided by applicant): Genetic mutations in genes encoding presenilins (PS1 and PS2) are responsible for the majority of early-onset familial AIzheimer's disease (FAD). Extensive studies in the past few years focusing on the mechanisms by which mutant PS promote AD pathogenesis have demonstrated that PS1 is a key component of gamma-secretase complex, an unusual protease catalyzing the intramembrane cleavage of amyloid precursor protein (APP) and generating pathogenic small peptide Abeta. Nascent full length PS1 polypeptides undergo endoproteolytic cleavage to produce N-terminal and C-terminal fragments (NTF and CTF) which associate with each other to form functional heterodimer. Early studies about PS1 metabolism suggest that unknown cellular factors tightly regulate biogenesis of PS1 heterodimer. However, identity of these limiting factors as well as other members of gamma-secretase complex has remained elusive. Through biochemical and genetic approaches, recent discoveries of three essential proteins (Nct, PEN-2 and APH-1) for gamma-secretase activity, provide us candidates for those hypothesized factors. Indeed, three of them have been demonstrated to be components of high molecular weight gamma-secretase complex, while their precise functions are unclear. My preliminary results demonstrated the regulatory roles of PEN-2 and APH-1 in PS1 cleavage and Nct maturation. This proposal is designed to further investigate the biological functions of PEN-2 and APH-1 in regulating PS1/gamma-secretase complex formation via examining PEN-2 and APH-1 structure/function relationship (Specific Aim 1), their impact on protein trafficking (Specific Aim 2), as well as protein folding and intermolecular interactions (Specific Aim 3) of PS1 and Nct.

描述（由申请人提供）：编码presenilins的基因（PS1和PS2）的基因突变负责大多数早期发作的家族性Aizheimer病（FAD）。在过去的几年中，广泛的研究着重于突变体PS促进AD发病机理的机制，这表明PS1是γ-分泌酶复合酶复合物的关键组成部分，γ-分泌酶复合物是一种催化淀粉样蛋白酶内膜前蛋白（APP）和发电pationic小肽Abeta的异常蛋白酶。新生的全长PS1多肽经历内蛋白溶解裂解，产生N末端和C末端片段（NTF和CTF），它们相互关联以形成功能异二聚体。关于PS1代谢的早期研究表明，未知的细胞因子严格调节PS1异二聚体的生物发生。但是，这些限制因素以及伽马分泌酶复合物的其他成员的身份仍然难以捉摸。通过生化和遗传学方法，对于伽马分泌酶活性的三种必需蛋白（NCT，PEN-2和APH-1）的最新发现为这些假设的因素提供了候选者。实际上，其中三个已被证明是高分子量γ-分泌酶复合酶复合酶的组成部分，而它们的精确功能尚不清楚。我的初步结果证明了PEN-2和APH-1在PS1裂解和NCT成熟中的调节作用。该建议旨在进一步研究PEN-2和APH-1在调节PS1/GAMMA-分泌酶复合物中的生物学功能，通过检查PEN-2和APH-1结构/功能关系（特定目标1），它们对蛋白质运输的影响（特定AIM 2），以及蛋白质折叠和蛋白质折叠和分子间相互作用（特定的AIM AIM 3）PS1和NCT和Nct anct and c1和nct anct ancc1和nct。