title: Mitochondrial stress causes increased succination of proteins in adipocytes in response to glucotoxicity.
abstract: 2SC [S-(2-succino)-cysteine] is a chemical modification formed by a Michael addition reaction of fumarate with cysteine residues in proteins. Formation of 2SC, termed 'succination' of proteins, increases in adipocytes grown in high-glucose medium and in adipose tissues of Type 2 diabetic mice. However, the metabolic mechanisms leading to increased fumarate and succination of protein in the adipocyte are unknown. Treatment of 3T3 cells with high glucose (30 mM compared with 5 mM) caused a significant increase in cellular ATP/ADP, NADH/NAD+ and Δψm (mitochondrial membrane potential). There was also a significant increase in the cellular fumarate concentration and succination of proteins, which may be attributed to the increase in NADH/NAD+ and subsequent inhibition of tricarboxylic acid cycle NAD+-dependent dehydrogenases. Chemical uncouplers, which dissipated Δψm and reduced the NADH/NAD+ ratio, also decreased the fumarate concentration and protein succination. High glucose plus metformin, an inhibitor of complex I in the electron transport chain, caused an increase in fumarate and succination of protein. Thus excess fuel supply (glucotoxicity) appears to create a pseudohypoxic environment (high NADH/NAD+ without hypoxia), which drives the increase in succination of protein. We propose that increased succination of proteins is an early marker of glucotoxicity and mitochondrial stress in adipose tissue in diabetes.
authors.affiliation: Department of Pharmacology, Physiology and Neuroscience, School of Medicine, University of South Carolina, Columbia, SC 29208, USA. norma.frizzell@uscmed.sc.edu
authors.forename: Norma
authors.initials: N
authors.lastname: Frizzell
authors.forename: Sonia A
authors.initials: SA
authors.lastname: Thomas
authors.forename: James A
authors.initials: JA
authors.lastname: Carson
authors.forename: John W
authors.initials: JW
authors.lastname: Baynes
mesh_terms: D016475:3T3 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival; D002952:Citric Acid Cycle; D015180:Electrophoresis, Gel, Two-Dimensional; D005650:Fumarates / Q000378:metabolism; D005947:Glucose / Q000633:toxicity*; D000860:Hypoxia; D008293:Malates / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D010085:Oxidative Phosphorylation; D010086:Oxidative Phosphorylation Coupling Factors; D018384:Oxidative Stress; D019802:Succinic Acid / Q000378:metabolism; D013549:Sweetening Agents / Q000633:toxicity*
chemical_list: D005650:Fumarates; D008293:Malates; D010086:Oxidative Phosphorylation Coupling Factors; D013549:Sweetening Agents; C030298:malic acid; D019802:Succinic Acid; D005947:Glucose