Development and Characterization of Quantitative, High-Throughput-Compatible Assays for Proteolytic Degradation of Glucagon.

ABSTRACT: Glucagon is a vital peptide hormone involved in the regulation of blood sugar under fasting conditions. Although the processes underlying glucagon production and secretion are well understood, far less is known about its degradation, which could conceivably be manipulated pharmacologically for therapeutic benefit. We describe here the development of novel assays for glucagon degradation, based on fluoresceinated and biotinylated glucagon (FBG) labeled at the N- and C-termini, respectively. Proteolysis at any peptide bond within FBG separates the fluorescent label from the biotin tag, which can be quantified in multiple ways. In one method requiring no specialized equipment, intact FBG is separated from the cleaved fluoresceinated fragments using NeutrAvidin agarose beads, and hydrolysis is

SUBMITTER: Suire CN 

PROVIDER: S-EPMC6585941 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature