Chemistry
Document Type
Article
Abstract
Insulin-degrading enzyme (IDE) is a zinc metalloprotease that selectively degrades biologically important substrates associated with type 2 diabetes and Alzheimer's disease (AD). As such, IDE is an attractive target for therapeutic innovations. Amajor requirement is an understanding of how other molecules present in cells regulate the activity of the enzyme toward insulin, IDE's most important physiologically relevant substrate. Previous kinetic studies of the IDE-dependent degradation of insulin in the presence of potential regulators have used iodinated insulin, a chemical modification that has been shown to alter the biological and biochemical properties of insulin. Here, we present a novel kinetic assay that takes advantage of the loss of helical circular dichroic signals of insulin with IDE-dependent degradation. As proof of concept, the resulting Michaelis-Menten kinetic constants accurately predict the known regulation of IDE by adenosine triphosphate (ATP). Intriguingly, we found that when Mg2+ is present with ATP, the regulation is abolished. The implication of this result for the development of preventative and therapeutic strategies for AD is discussed. We anticipate that the new assay presented here will lead to the identification of other small molecules that regulate the activity of IDE toward insulin.
Publication Title
Bioscience Reports
Publication Date
11-7-2018
Volume
38
Issue
6
ISSN
0144-8463
DOI
10.1042/BSR20181416
Repository Citation
Ivancic, Valerie A.; Krasinski, Claire A.; Zheng, Qiuchen; Meservier, Rebecca J.; Spratt, Donald E.; and Lazo, Noel D., "Enzyme kinetics from circular dichroism of insulin reveals mechanistic insights into the regulation of insulin-degrading enzyme" (2018). Chemistry. 126.
https://commons.clarku.edu/chemistry/126
Cross Post Location
Student Publications
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyright Conditions
Ivancic, V. A., Krasinski, C. A., Zheng, Q., Meservier, R. J., Spratt, D. E., & Lazo, N. D. (2018). Enzyme kinetics from circular dichroism of insulin reveals mechanistic insights into the regulation of insulin-degrading enzyme. Bioscience reports, 38(6), BSR20181416.
