title: Redox regulation of insulin degradation by insulin-degrading enzyme.
abstract: Insulin-degrading enzyme (IDE) is a thiol sensitive peptidase that degrades insulin and amyloid β, and has been linked to type 2 diabetes mellitus and Alzheimer's disease. We examined the thiol sensitivity of IDE using S-nitrosoglutathione, reduced glutathione, and oxidized glutathione to distinguish the effects of nitric oxide from that of the redox state. The in vitro activity of IDE was studied using either partially purified cytosolic enzyme from male Sprague-Dawley rats, or purified rat recombinant enzyme. We confirm that nitric oxide inhibits the degrading activity of IDE, and that it affects proteasome activity through this interaction with IDE, but does not affect the proteasome directly. Oxidized glutathione inhibits IDE through glutathionylation, which was reversible by dithiothreitol but not by ascorbic acid. Reduced glutathione had no effect on IDE, but reacted with partially degraded insulin to disrupt its disulfide bonds and accelerate its breakdown to trichloroacetic acid soluble fragments. Our results demonstrate the sensitivity of insulin degradation by IDE to the redox environment and suggest another mechanism by which the cell's oxidation state may contribute to the development of, and the link between, type 2 diabetes and Alzheimer's disease.
authors: Research Service, Omaha Veterans Affairs Medical Center, Omaha, Nebraska, United States of America.
authors: Crystal M
authors: CM
authors: Cordes
authors: Robert G
authors: RG
authors: Bennett
authors: Gerri L
authors: GL
authors: Siford
authors: Frederick G
authors: FG
authors: Hamel
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D019803:Glutathione Disulfide / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D007339:Insulysin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008297:Male; D015538:Nitrosation / Q000187:drug effects; D010084:Oxidation-Reduction / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D026422:S-Nitrosoglutathione / Q000494:pharmacology
chemical_list: D016229:Amyloid beta-Peptides; D007328:Insulin; D026422:S-Nitrosoglutathione; D002918:Chymotrypsin; D007339:Insulysin; D046988:Proteasome Endopeptidase Complex; D004229:Dithiothreitol; D019803:Glutathione Disulfide