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Yeast Ste23p Shares Functional Similarities With Mammalian Insulin-Degrading Enzymes, Benjamin Alper, Jarrad W. Rowse, Walter K. Schmidt
Yeast Ste23p Shares Functional Similarities With Mammalian Insulin-Degrading Enzymes, Benjamin Alper, Jarrad W. Rowse, Walter K. Schmidt
Chemistry & Physics Faculty Publications
The S. cerevisiae genome encodes two M16A enzymes: Axl1p and Ste23p. Of the two, Ste23p shares significantly higher sequence identity with M16A enzymes from other species, including mammalian insulin-degrading enzymes (IDEs). In this study, recombinant Ste23p and R. norvegicus IDE (RnIDE) were isolated from E. coli, and their enzymatic properties compared. Ste23p was found to cleave established RnIDE substrates including the amyloid-β peptide (Aβ 1-40) and insulin B chain. A novel internally quenched fluorogenic substrate (Abz-SEKKDNYIIKGVnitroY-OH) based on the polypeptide sequence of the yeast P2 a-factor mating propheromone was determined to be a suitable substrate for …
A Capillary Electrophoresis Method For Evaluation Of Abeta Proteolysis In Vitro, Benjamin J. Alper, Walter K. Schmidt
A Capillary Electrophoresis Method For Evaluation Of Abeta Proteolysis In Vitro, Benjamin J. Alper, Walter K. Schmidt
Chemistry & Physics Faculty Publications
According to the amyloid hypothesis, Aβ peptides are neurotoxic and underlie development and progression of Alzheimer’s disease (AD). Multiple Aβ clearance mechanisms, including destruction of the peptides by proteolytic enzymes, are hypothesized to regulate physiological Aβ peptide levels. The insulin-degrading enzyme (IDE) is considered one of the predominant enzymes having Aβ 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 Aβ 1–40 proteolysis. This method employs chemically unmodified substrates that are readily obtained …