γ-分泌酶/底物相互作用的单分子动力学研究以及 AD 引起的突变的影响-猫眼课题宝

γ-分泌酶/底物相互作用的单分子动力学研究以及 AD 引起的突变的影响

批准号：

10323672

负责人：

Scott Thomas Forth

金额：

$19.27万

依托单位：

RENSSELAER POLYTECHNIC INSTITUTE

依托单位国家：

美国

项目类别：

财政年份：

2021

资助国家：

美国

项目状态：

已结题

起止时间：

2021-01-15 至 2023-12-31

来源：

https://reporter.nih.gov/project-details/10323672

关键词：

Active Sites Alzheimer&apos s Disease Amyloid Amyloid beta-Protein Amyloid beta-Protein Precursor Aspartate Benign Binding Biochemical Biological Assay Brain C-terminal Carboxypeptidase Catalytic Domain Complex Coupled Dementia Docking Dyes Enzyme Interaction Enzyme Kinetics Enzymes Experimental Designs Fluorescence Generations Homologous Gene Immobilization Individual Integral Membrane Protein Kinetics Lipid Bilayers Liposomes Measurement Measures Mediating Membrane Methods Micelles Microscope Monitor Mutation Pathologic Peptide Fragments Peptide Hydrolases Process Production Proteolysis Public Health Senile Plaques Substrate Interaction Techniques Time Total Internal Reflection Fluorescent Transmembrane Domain United States beta pleated sheet beta secretase disease-causing mutation drug discovery early onset enzyme substrate familial Alzheimer disease gamma secretase hydrophilicity insight mutant neurotoxic novel presenilin protein complex single molecule

中文摘要

翻译

项目摘要淀粉样斑块是阿尔茨海默病的病理标志，由淀粉样蛋白-b多肽(A-b)组成疾病(AD)。G分泌酶负责C99的裂解，C99是99的C末端片段淀粉样前体蛋白(APP)的残基，产生抗体。前人对g-的动力学研究分泌酶测定APP胞内区(AICD)和/或抗体的最终产量。然而，从C99产生抗体的各个步骤的动力学速率是缺乏的。在这里，我们将使用单分子荧光研究来观察酶/底物分子实时测定酶/底物结合和裂解动力学。分泌酶介导的膜内蛋白分解产生抗体(Aim1)，并确定如何家族性AD(FAD)突变改变酶/底物结合和切割动力学 AD(AIM2)。

英文摘要

Project Summary Amyloid plaque, composed of amyloid-b peptide (Ab), is a pathological hallmark of Alzheimer’s disease (AD). g-secretase is responsible for the cleavage of C99, the C-terminal fragment of 99 residues of amyloid precursor protein (APP), to generate Ab. Previous kinetics studies of g- secretase measured the final production of APP intracellular domain (AICD) and/or Ab. However, the kinetics rates for individual steps of the generation of Ab from C99 are lacking. Here we will use single molecule fluorescence studies to observe enzyme/substrate molecules in real time, and measure the kinetics of enzyme/substrate association and cleavage in g- secretase-mediated intramembrane proteolysis to generate Ab (Aim1), and determine how familial AD (FAD) mutations alter the kinetics of enzyme/substrate association and cleavage in AD (Aim2).

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