Uncovering Protein Interactions and Membrane Phase Preferences that alter the Plasma Membrane trafficking of Peripheral Myelin Protein 22

揭示改变外周髓鞘蛋白质膜运输的蛋白质相互作用和膜相偏好 22

• 批准号: 10011580
• 负责人: Justin Tyler Marinko
• 金额: $ 1.59万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2020-12-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011580
• 关键词: Actins; Address; Adhesions; Affect; Alzheimer' s Disease; Binding; Biological; Biological Assay; Cell membrane; Cell physiology; Cell surface; Cells; Charcot-Marie-Tooth Disease; Cholesterol; Collaborations; Confocal Microscopy; Crowding; Cytoskeleton; DNA Sequence Alteration; Data; Disease; Endoplasmic Reticulum; Environment; Flow Cytometry; Gene Deletion; Gene Dosage; Gene Duplication; Genes; Goals; Hela Cells; Hereditary neuropathy with liability to pressure palsies; Hydrophobicity; Impairment; Individual; Integral Membrane Protein; Lead; Link; Mass Spectrum Analysis; Mediating; Membrane; Membrane Proteins; Microscopy; Missense Mutation; Molecular; Molecular Conformation; Monitor; Mus; Mutate; Mutation; Mutation Analysis; Myelin; Neurodegenerative Disorders; Neuropathy; PMP22 gene; Parkinson Disease; Patients; Peripheral Nervous System; Peripheral Nervous System Diseases; Phase; Play; Post-Translational Protein Processing; Process; Protein Glycosylation; Proteins; Proteomics; Quality Control; Rattus; Research; Research Personnel; Role; Schwann Cells; Scientist; Small Interfering RNA; Syndrome; System; Testing; Therapeutic; Transfection; Vesicle; Work; base; career; human disease; knock-down; member; migration; mutant; novel; novel therapeutics; overexpression; palmitoylation; preference; protein expression; protein folding; screening; small molecule; small molecule libraries; symptom management; trafficking

PROJECT SUMMARY: A major goal of my career as an independent researcher is to understand how integral membrane proteins fold in the crowded environment of the endoplasmic reticulum (ER). Disruptions in the fidelity of this process lead to a number of human diseases including Alzheimer’s, Parkinson’s and Charcot-Marie-Tooth Disease (CMTD). By better understanding this process, I hope to eventually identify novel therapeutic strategies for treating patients afflicted with these diseases. CMTD is the most common neuropathy of the peripheral nervous system (PNS) and afflicts 1 in 2,500 individuals. Over 80% of patients afflicted with this disease have a mutation in the protein encoding PMP22 gene. PMP22 is a tetraspan integral membrane protein that is highly expressed at the plasma membrane (PM) of myelinating Schwann cells of the PNS. Improper gene dosage of PMP22 in CMTD patients results in either too much or too little PMP22 at the PM of Schwann cells. The altered amount of PMP22 at the PM is believed to cause the myelin abnormalities observed in these patients. There are currently no treatments for CMTD. PMP22 folds in the ER under the surveillance of the ER quality control (QC) system. This system both helps PMP22 to fold as well as mediates the ultimate decision whether to traffic PMP22 forward towards the PM or to retain the protein in the ER and ultimately target it for degradation. The goals of this proposal are to understand the factors that mediate this decision process. Previous results show that wild type (WT) PMP22 and PMP22 disease mutants have different protein interactions in the ER QC system leading to varying PM expression. Preliminary data shows that selective modulation of protein expression in ER can alter the cell surface trafficking of PMP22 and its disease mutants. Additionally, PMP22 associates with cholesterol rich membrane domains at the PM. It has been shown for another membrane protein of the PNS, P0, that association with cholesterol in the ER is necessary for cell surface trafficking. Schwann cells derived from PMP22 -/- mice show decreased migration and adhesion capabilities and lower PM cholesterol content compared to WT Schwann cells. This indicates that PMP22 association with cholesterol rich domains is important for Schwann cell function. It is my hypothesis that PMP22 cell surface trafficking can be modulated by both changing its protein interaction network in the ER and through altering its ability to associate with cholesterol rich membrane domains. To address this hypothesis, I will: (1) uncover mutation specific changes to the PMP22 interactome in the ER that modulate its cell surface trafficking and (2) examine the factors that cause PMP22 to associate with cholesterol rich membrane domains and assess the effects of cholesterol association on PMP22 trafficking. The results from this proposal will uncover novel protein interactions and factors that affect PMP22 trafficking and cholesterol association. This information will open novel avenues for scientists attempting to develop therapeutics for patients who suffer from CMTD.

项目摘要：我作为独立研究人员的我护理护理职业的主要目标是了解不可或缺的 膜蛋白在内质网（ER）拥挤的环境中折叠。 过程导致许多人类疾病，包括阿尔茨海默氏症，帕金森氏症和charcot-marcot-marcot齿病 （CMTD）。 患有这些疾病的患者。 折磨2500人中有80％的人在蛋白质编码中具有突变 PMP22基因。 PNS的骨髓细胞不当。 Schwann细胞PM的PMP22太少。 这些患者目前尚无CMTD治疗。 ER质量控制（QC）系统的监视。 最终决定是将PMP22转移到PM还是ER中的蛋白质并最终靶向蛋白质的最终决定 它的降级是调解该决策过程的因素。 结果表明，野生型（WT）PMP22和PMP22疾病突变体在EER QC中具有不同的蛋白质相互作用 导致PM表达不同的系统。 可以改变PMP22的细胞表面运输及其疾病杂物。 PM的丰富膜域已显示为PNS的另一个膜 ER中的胆固醇是细胞表面运输所必需的。 与WT Schwann细胞相比，迁移和粘附能力降低和PM胆固醇含量降低。 表明PMP22与富含胆固醇的富域的关联对于Schwann细胞功能很重要。 假设PMP22细胞表面运输可以通过改变其蛋白质相互作用网络的模块化。 通过改变其与富含胆固醇的膜结合的能力。 威尔：（1）在ER中，pmp22相互作用的特定变化特异性变化该模块细胞表面运输 （2）检查导致PMP22与富含胆固醇的膜结构域关联的因素并评估效果 PMP22运输的胆固醇协会。 影响PMP22贩运和胆固醇协会的因素。 试图为患有CMTD的患者开发治疗疗法。