Structure, Folding, and Misfolding of PMP22

PMP22 的结构、折叠和错误折叠

• 批准号: 8247008
• 负责人: Bruce D Carter
• 金额: $ 30.58万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247008
• 关键词: Afferent Neurons; Amino Acids; Biological; Cell Line; Cell surface; Cells; Charcot-Marie-Tooth Disease; Chemicals; Coculture Techniques; Defect; Disease; Genes; Goals; Human; Human Cell Line; Kinetics; Lead; Link; Maintenance; Membrane Proteins; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Mutation; Myelin; Myelin Proteins; NMR Spectroscopy; Nature; Pathway interactions; Peripheral; Peripheral Nervous System Diseases; Production; Proteins; Research; Role; Schwann Cells; Structure; Structure-Activity Relationship; Testing; Transgenic Mice; Work; base; cytotoxic; human PMP22 protein; human disease; insight; mouse model; mutant; novel therapeutics; overexpression; protein function; protein structure; trafficking

Project Summary Genetically dominant mutations in the gene that encodes peripheral myelin protein 22 (PMP22) lead to single amino acid changes in its sequence that result in defective myelin, underlying the common human peripheral neuropathy, Charcot-Marie-Tooth Disease Type IA (CMTD). It is believed that CMTD mutations result in misassembly of PMP22 early in the secretory pathway, resulting in the loss of protein function and also in the formation of potentially cytotoxic aggregates. The overall goal of this project is to elucidate the molecular biophysical nature of the perturbations made by CMTD-associated mutations to the structure, stability and folding of this critical membrane protein. We also seek to test whether chemical chaperones can correct the folding defects normally observed for CMTD mutant forms of PMP22. Aim 1. Characterize the structures of the wild type (WT) and CMTD mutant forms of human PMP22 using NMR spectroscopy. Aim 1 will test the hypothesis that CMTD mutant forms of PMP22 differ from the WT protein in terms of conformation and/or oligomeric state. Structural information will also illuminate PMP22's structure/function relationships and its role in myelin production and maintenance, and will provide biophysical insight into how amino acid mutations result in CMTD. Determination of PMP22's structure will also add to the currently sparse gallery of human membrane protein structures. Aim 2. Characterize the stability and folding kinetics of WT and CMTD mutant forms of PMP22. Aim 2 will test the hypothesis that disease-related mutations of PMP22 destabilize the protein. Aim 2 will also test the hypothesis that disease-related mutant forms of PMP22 fold more slowly and/or inefficiently than the wild type protein. We will also test whether CMTD mutant forms of PMP22 are aggregation-prone. Aim 3. Determine whether chemical chaperones can increase the cell surface expression of CMTD mutant forms of PMP22. Aim 3 will test the hypothesis that PMP22 is akin to other human membrane proteins that are linked to diseases involving protein misassembly and for which it has already been established that proper folding and trafficking can be restored using chemical chaperones.

项目摘要 编码外周髓鞘蛋白22(PMP22)基因的遗传显性突变 导致ITS序列中单一氨基酸的变化，导致髓鞘缺陷，从而导致 常见的人类周围神经病，夏科-玛丽-图斯病I型(CMTD)。它是 认为CMTD突变导致PMP22在分泌途径早期错误组装， 导致蛋白质功能丧失，并形成潜在的细胞毒性 集合体。这个项目的总体目标是阐明人类免疫缺陷病毒的分子生物物理性质。 CMTD相关突变对细胞结构、稳定性和折叠的影响 这种关键的膜蛋白。我们还试图测试化学伴侣是否可以纠正 通常在CMTD突变形式的PMP22中观察到的折叠缺陷。 目的1.鉴定野生型(WT)和CMTD突变体的结构 人PMP22的核磁共振波谱分析。目标1将检验CMTD突变的假设 PMP22的不同形式与WT蛋白在构象和/或低聚状态方面不同。 结构信息还将阐明PMP22的S结构/功能关系及其在 髓鞘的产生和维护，并将提供生物物理洞察如何氨基酸 突变会导致CMTD。PMP22的S结构的确定也将增加目前的 人膜蛋白结构的稀疏图库。 目的2.鉴定WT和CMTD突变体的稳定性和折叠动力学 PMP22。目的2将检验与疾病相关的PMP22突变会破坏 蛋白。Aim 2还将测试与疾病相关的PMP22突变形式折叠的假设 比野生型蛋白更慢和/或更低效。我们还将测试CMTD是否 PMP22的突变形式易于聚集。 目的3.确定化学伴侣是否能增加细胞表面的表达 PMP22的CMTD突变形式。目标3将检验PMP22与其他基因相似的假设 与涉及蛋白质错误组装的疾病有关的人膜蛋白 它已经被确定可以用来恢复正确的折叠和贩运 化学伴侣。