Degradation of amylin by insulin-degrading enzyme.
A pathological feature of Type 2 diabetes is deposits in the pancreatic islets primarily composed of amylin (islet amyloid polypeptide). Although much attention has been paid to the expression and secretion of amylin, little is known about the enzymes involved in amylin turnover. Recent reports suggest that insulin-degrading enzyme (IDE) may have specificity for amyloidogenic proteins, and therefore we sought to determine whether amylin is an IDE substrate. Amylin-degrading activity co-purified with IDE from rat muscle through several chromatographic steps. Metalloproteinase inhibitors inactivated amylin-degrading activity with a pattern consistent with the enzymatic properties of IDE, whereas inhibitors of acid and serine proteases, calpains, and the proteasome were ineffective. Amylin degradation was inhibited by insulin in a dose-dependent manner, whereas insulin degradation was inhibited by amylin. Other substrates of IDE such as atrial natriuretic peptide and glucagon also competitively inhibited amylin degradation. Radiolabeled amylin and insulin were both covalently cross-linked to a protein of 110 kDa, and the binding was competitively inhibited by either unlabeled insulin or amylin. Finally, a monoclonal anti-IDE antibody immunoprecipitated both insulin- and amylin-degrading activities. The data strongly suggest that IDE is an amylin-degrading enzyme and plays an important role in the clearance of amylin and the prevention of islet amyloid formation.
Bennett
R G
RG
Medical Research Service, Omaha Veterans Affairs Medical Center and Department of Internal Medicine, University of Nebraska Medical Center, Omaha, Nebraska 68105, USA. rgbennet@unmc.edu
Duckworth
W C
WC
Hamel
F G
FG
D000682:Amyloid / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007339:Insulysin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D058228:Islet Amyloid Polypeptide; D008297:Male; D018482:Muscle, Skeletal / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000682:Amyloid; D007328:Insulin; D058228:Islet Amyloid Polypeptide; D007339:Insulysin