title: Carbonylation contributes to SERCA2a activity loss and diastolic dysfunction in a rat model of type 1 diabetes.
abstract: OBJECTIVE
  Approximately 25% of children and adolescents with type 1 diabetes will develop diastolic dysfunction. This defect, which is characterized by an increase in time to cardiac relaxation, results in part from a reduction in the activity of the sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), the ATP-driven pump that translocates Ca(2+) from the cytoplasm to the lumen of the sarcoplasmic reticulum. To date, mechanisms responsible for SERCA2a activity loss remain incompletely characterized.
  
  
  METHODS
  The streptozotocin (STZ)-induced murine model of type 1 diabetes, in combination with echocardiography, high-speed video detection, confocal microscopy, ATPase and Ca(2+) uptake assays, Western blots, mass spectrometry, and site-directed mutagenesis, were used to assess whether modification by reactive carbonyl species (RCS) contributes to SERCA2a activity loss.
  
  
  RESULTS
  After 6-7 weeks of diabetes, cardiac and myocyte relaxation times were prolonged. Total ventricular SERCA2a protein remained unchanged, but its ability to hydrolyze ATP and transport Ca(2+) was significantly reduced. Western blots and mass spectroscopic analyses revealed carbonyl adducts on select basic residues of SERCA2a. Mutating affected residues to mimic physio-chemical changes induced on them by RCS reduced SERCA2a activity. Preincubating with the RCS, methylglyoxal (MGO) likewise reduced SERCA2a activity. Mutating an impacted residue to chemically inert glutamine did not alter SERCA2a activity, but it blunted MGO's effect. Treating STZ-induced diabetic animals with the RCS scavenger, pyridoxamine, blunted SERCA2a activity loss and minimized diastolic dysfunction.
  
  
  CONCLUSIONS
  These data identify carbonylation as a novel mechanism that contributes to SERCA2a activity loss and diastolic dysfunction during type 1 diabetes.
authors.affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Chun Hong
authors.initials: CH
authors.lastname: Shao
authors.forename: Haley L
authors.initials: HL
authors.lastname: Capek
authors.forename: Kaushik P
authors.initials: KP
authors.lastname: Patel
authors.forename: Mu
authors.initials: M
authors.lastname: Wang
authors.forename: Kang
authors.initials: K
authors.lastname: Tang
authors.forename: Cyrus
authors.initials: C
authors.lastname: DeSouza
authors.forename: Ryoji
authors.initials: R
authors.lastname: Nagai
authors.forename: William
authors.initials: W
authors.lastname: Mayhan
authors.forename: Muthu
authors.initials: M
authors.lastname: Periasamy
authors.forename: Keshore R
authors.initials: KR
authors.lastname: Bidasee
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism* / Q000503:physiopathology; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism* / Q000503:physiopathology; D003971:Diastole / Q000502:physiology*; D006321:Heart / Q000503:physiopathology*; D008297:Male; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D009206:Myocardium / Q000378:metabolism*; D050050:Protein Carbonylation; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*
chemical_list: D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases