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Conformational and functional analysis of Apolipoprotein E

载脂蛋白E的构象和功能分析

基本信息

批准号：
10334412
负责人：
Andrea Soranno
金额：
$ 44.45万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10334412
关键词：
Adopted Affinity Alleles Alzheimer&apos s Disease Alzheimer&apos s disease risk Apolipoprotein E Binding Binding Proteins Biochemical Cardiovascular Pathology Cardiovascular system Carrier Proteins Cerebrospinal Fluid Cholesterol Homeostasis Complex Data Disease Equilibrium Exhibits Fluorescence Resonance Energy Transfer Fluorescence Spectroscopy Health Heterogeneity Late Onset Alzheimer Disease Length Ligands Light Link Lipid Binding Lipids Lipoproteins Methods Modeling Molecular Molecular Conformation Movement Pathogenicity Physiological Plasma Play Point Mutation Property Protein Conformation Protein Fragment Protein Isoforms Proteins Risk Role Structural Models Structure Testing Thinking Time Toxic effect Variant Vesicle apolipoprotein E-3 base cardiovascular disorder risk conformer experimental study flexibility genetic risk factor interest lipid transport monomer nanodisk nanosecond novel novel strategies novel therapeutic intervention particle protein function recruit single molecule single-molecule FRET small molecule structural biology

项目摘要

ABSTRACT The ε4-allele isoform of apolipoprotein E (ApoE4) plays a key-role in Alzheimer's disease and cardiovascular pathologies. A large body of evidence support that conformations of the protein are instrumental in its contribution to function and disease; yet, much remains unknown about the conformational ensemble of full-length ApoE and its role in protein (dis)function, largely because of its elevated propensity for aggregation/oligomerization and inherent flexibility. In our lab, we have overcome these complications by harnessing state-of-the-art single-molecule fluorescence spectroscopy and, for the first time, we are able to access the structural ensemble of the monomeric full-length ApoE4 (free in solution, embedded in oligomers, and bound to lipids). Our preliminary data clearly indicate that ApoE4 adopts at least three distinct conformers – previously unidentified – that coexist in equilibrium. These conformers are highly dynamic and malleable to oligomerization and lipid binding. These novel observations led us to hypothesize that single-point mutations and small molecules can perturb the structural ensemble favoring/disfavoring specific “pathogenic” conformations, whereas interaction with ligands (e.g. lipids) alters the equilibrium between the different conformers selecting for specific conformations that are required for protein function. Our approach will allow us to determine the conformational properties of ApoE, probe current structural models, and test our hypothesis. The specific aims are as follows: 1) determine the conformational changes and domain movements within the pathogenic ApoE4 and elucidate how point mutations and small molecules modulate protein toxicity; 2) understand the mechanism of lipid interactions, from synthetic lipid vesicles to physiological lipoproteins. A comprehensive description of the structural conformations and dynamics of ApoE in the presence and absence of lipids will shed light on the molecular mechanism behind its role in health and disease, paving the way to developing new therapeutic strategies.

摘要