A capillary electrophoresis method for evaluation of Aβ proteolysis in vitro

被引：14

作者：

Alper, Benjamin J. ^{[1
]}

Schmidt, Walter K. ^{[1
]}

机构：

[1] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2009年 / 178卷 / 01期

关键词：

Alzheimer's disease; Amyloid beta; Insulin-degrading enzyme; Capillary electrophoresis; INSULIN-DEGRADING ENZYME; AMYLOID PRECURSOR PROTEIN; ALZHEIMERS-DISEASE; EXTRACELLULAR LEVELS; PEPTIDES; DEGRADATION; PLAQUES; BRAIN; RISK; GENE;

D O I：

10.1016/j.jneumeth.2008.11.010

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

According to the amyloid hypothesis, A beta peptides are neurotoxic and underlie development and progression of Alzheimer's disease (AD). Multiple A beta clearance mechanisms, including destruction of the peptides by proteolytic enzymes, are hypothesized to regulate physiological A beta peptide levels. The insulin-degrading enzyme (IDE) is considered one of the predominant enzymes having A beta degrading activity. Despite its putative role in protecting against AD, relatively few methods exist for studying IDE activity in vitro. We report the application of capillary electrophoresis (CE) as a novel method for evaluating IDE-mediated A beta 1-40 proteolysis. This method employs chemically unmodified substrates that are readily obtained from commercial sources. It involves minimal sample preparation, and requires no specialized equipment beyond a CE instrument equipped with a standard fused silica capillary. In the present analysis, we demonstrate that this CE-based method is amenable to kinetic analysis, and show that IDE-mediated A beta proteolysis is significantly and disproportionately inhibited in the presence of insulin, an alternative IDE substrate. Published by Elsevier B.V.

引用

页码：40 / 45

页数：6

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