Article
Ghrelin Proteolysis by Insulin-degrading Enzyme
bioRxiv
(2021)
Abstract
Here we report proteolysis of synthetic acylated human ghrelin by recombinant human insulin-degrading enzyme (IDE). Kinetic parameters and sites of proteolytic cleavage were evaluated. Ghrelin proteolysis by IDE was inhibited by ethylenediaminetetraacetate (EDTA), a metal chelating agent. Ghrelin proteolysis appears at least somewhat specific to M16 family proteases such as IDE, as the M13 protease neprilysin (NEP) did not exhibit ghrelin proteolysis in this study. A quenched fluorogenic peptide substrate comprising the primary sites of IDE-mediated ghrelin proteolysis (McaQRVQQRKESKK(Dnp)-OH; Mca: 7-methoxycoumarin-3-carboxylic acid; Dnp: 2,4-dinitrophenyl) was developed and used to evaluate enzyme specificity and kinetic parameters of proteolysis. Like acyl ghrelin, Mca-QRVQQRKESKK(Dnp)-OH was efficiently cleaved by IDE central to the target sequence. We anticipate that this quenched fluorogenic peptide substrate will be of value to future studies of ghrelin proteolysis by IDE and potentially other peptidases.
Disciplines
Publication Date
June, 2021
DOI
10.1101/2021.06.10.447714
Publisher Statement
bioRxiv preprint doi: https://doi.org/10.1101/2021.06.10.447714; this version posted June 10, 2021. The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Link to publication: https://www.biorxiv.org/content/10.1101/2021.06.10.447714v1
Citation Information
David D. Bocach, Kierstin L. Jones, Jonathan M. Bell, Qiuchen Zheng, et al.. "Ghrelin Proteolysis by Insulin-degrading Enzyme" bioRxiv (2021) Available at: http://works.bepress.com/benjamin_alper/28/