title: Redox proteomics identification of oxidatively modified myocardial proteins in human heart failure: implications for protein function.
abstract: Increased oxidative stress in a failing heart may contribute to the pathogenesis of heart failure (HF). The aim of this study was to identify the oxidised proteins in the myocardium of HF patients and analyse the consequences of oxidation on protein function. The carbonylated proteins in left ventricular tissue from failing (n = 14) and non-failing human hearts (n = 13) were measured by immunoassay and identified by proteomics. HL-1 cardiomyocytes were incubated in the presence of stimuli relevant for HF in order to assess the generation of reactive oxygen species (ROS), the induction of protein carbonylation, and its consequences on protein function. The levels of carbonylated proteins were significantly higher in the HF patients than in the controls (p<0.01). We identified two proteins that mainly underwent carbonylation: M-type creatine kinase (M-CK), whose activity is impaired, and, to a lesser extent, α-cardiac actin. Exposure of cardiomyocytes to angiotensin II and norepinephrine led to ROS generation and M-CK carbonylation with loss of its enzymatic activity. Our findings indicate that protein carbonylation is increased in the myocardium during HF and that these oxidative changes may help to explain the decreased CK activity and consequent defects in energy metabolism observed in HF.
authors.affiliation: Centro Cardiologico Monzino IRCCS, Milan, Italy.
authors.forename: Maura
authors.initials: M
authors.lastname: Brioschi
authors.forename: Gianluca
authors.initials: G
authors.lastname: Polvani
authors.forename: Pasquale
authors.initials: P
authors.lastname: Fratto
authors.forename: Alessandro
authors.initials: A
authors.lastname: Parolari
authors.forename: Piergiuseppe
authors.initials: P
authors.lastname: Agostoni
authors.forename: Elena
authors.initials: E
authors.lastname: Tremoli
authors.forename: Cristina
authors.initials: C
authors.lastname: Banfi
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D052283:Creatine Kinase, MM Form / Q000378:metabolism; D004734:Energy Metabolism; D005260:Female; D006333:Heart Failure / Q000378:metabolism* / Q000503:physiopathology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009124:Muscle Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology; D009206:Myocardium / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D050050:Protein Carbonylation; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D009124:Muscle Proteins; D017382:Reactive Oxygen Species; D052283:Creatine Kinase, MM Form