Chronic Diabetes Increases Advanced Glycation End Products on Cardiac Ryanodine Receptors/Calcium-Release Channels

• Published in: Diabetes (New York, N.Y.) Vol. 52; no. 7; pp. 1825 - 1836
• Main Authors: Bidasee, Keshore R, Nallani, Karuna, Yu, Yongqi, Cocklin, Ross R, Zhang, Yinong, Wang, Mu, Dincer, U Deniz, Besch, Jr, Henry R
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Diabetes Association 01.07.2003
• Subjects: Amino Acid Sequence; Animals; Binding Sites; Biological and medical sciences; Cardiomyopathy; Chemical bonds; Complications; Development and progression; Diabetes; Diabetes mellitus; Diabetes Mellitus, Experimental - genetics; Diabetes Mellitus, Experimental - metabolism; Diabetes Mellitus, Experimental - physiopathology; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation - physiology; Glucose; Glycation End Products, Advanced - metabolism; Heart - physiopathology; Heart diseases; Insulin; Male; Myocardial diseases; Myocardium - metabolism; Peptide Fragments; Peptides; Proteins; Rats; Rats, Sprague-Dawley; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; Ryanodine - metabolism; Ryanodine Receptor Calcium Release Channel - genetics; Ryanodine Receptor Calcium Release Channel - physiology; Trypsin; United States; United States > US; Indianapolis Indiana
• ISSN: 0012-1797; 1939-327X
• DOI: 10.2337/diabetes.52.7.1825

Chronic Diabetes Increases Advanced Glycation End Products on Cardiac Ryanodine Receptors/Calcium-Release Channels Keshore R. Bidasee 1 , Karuna Nallani 2 , Yongqi Yu 2 , Ross R. Cocklin 3 , Yinong Zhang 3 , Mu Wang 3 , Ü. Deniz Dincer 4 and Henry R. Besch, Jr. 2 5 1 Department of Pharmacology, University of Nebraska Medical Center, Omaha, Nebraska 2 Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana 3 Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana 4 Department of Pharmacology, Faculty of Pharmacy, University of Ankara, Tandogan, Ankara, Turkey 5 Krannert Institute of Cardiology, Center for Vascular Biology and Medicine, Indianapolis, Indiana Address correspondence and reprint requests to Keshore R. Bidasee, Department of Pharmacology, University of Nebraska Medical Center, 986260 Nebraska Medical Center, Omaha, NE 68198-6260. E-mail: kbidasee{at}unmc.edu Abstract Decrease in cardiac contractility is a hallmark of chronic diabetes. Previously we showed that this defect results, at least in part, from a dysfunction of the type 2 ryanodine receptor calcium-release channel (RyR2). The mechanism(s) underlying RyR2 dysfunction is not fully understood. The present study was designed to determine whether non-cross-linking advanced glycation end products (AGEs) on RyR2 increase with chronic diabetes and if formation of these post-translational complexes could be attenuated with insulin treatment. Overnight digestion of RyR2 from 8-week control animals (8C) with trypsin afforded 298 peptides with monoisotopic mass (M+H + ) ≥500. Digestion of RyR2 from 8-week streptozotocin-induced diabetic animals (8D) afforded 21% fewer peptides, whereas RyR2 from 6-week diabetic/2-week insulin-treated animals generated 304 peptides. Using an in-house PERLscript algorithm, search of matrix-assisted laser desorption ionization-time of flight mass data files identified several M+H + peaks corresponding to theoretical RyR2 peptides with single N ε -(carboxymethyl)-lysine, imidazolone A, imidazone B, pyrraline, or 1-alkyl-2-formyl-3,4-glycosyl pyrrole modification that were present in 8D but not 8C. Insulin treatment minimized production of some of these nonenzymatic glycation products. These data show for the first time that AGEs are formed on intracellular RyR2 during diabetes. Because AGE complexes are known to compromise protein activity, these data suggest a potential mechanism for diabetes-induced RyR2 dysfunction. 6D-2I, 6-week streptozotocin-induced diabetic/2-week insulin-treated animals 8C, 8-week control animals 8D, 8-week streptozotocin-induced diabetic animals AFGP, 1-alkyl-2-formyl-3,4-glycosyl pyrrole molecule AGE, advanced glycation end product Kd, equilibrium dissociation constant M+H+, monoisotopic mass MALDI-TOF, matrix-assisted laser desorption ionization-time of flight RAGE, receptor for AGEs RyR2, type 2 ryanodine receptor calcium-release channel STZ, streptozotocin Footnotes Accepted March 31, 2003. Received January 26, 2003. DIABETES