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Redox proteomics identification of oxidatively modified myocardial proteins in human heart failure: implications for protein function.

Brioschi M, Polvani G, Fratto P, Parolari A, Agostoni P, Tremoli E, Banfi C - PLoS ONE (2012)

Bottom Line: The levels of carbonylated proteins were significantly higher in the HF patients than in the controls (p<0.01).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.

View Article: PubMed Central - PubMed

Affiliation: Centro Cardiologico Monzino IRCCS, Milan, Italy.

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.

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Detection of protein carbonyls after immunoprecipitation of M-CK from cell lysate.HL-1 cells were treated with angiotensin II (AngII) or phenylephrine (PE) for 24 h, and the immunoprecipitates were immunoblotted with anti-DNP and anti-CK. Representative of three independent experiments.
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pone-0035841-g005: Detection of protein carbonyls after immunoprecipitation of M-CK from cell lysate.HL-1 cells were treated with angiotensin II (AngII) or phenylephrine (PE) for 24 h, and the immunoprecipitates were immunoblotted with anti-DNP and anti-CK. Representative of three independent experiments.

Mentions: In comparison with the controls, M-CK protein carbonylation was significantly increased by 2.5±0.9 fold in the angiotensin II-treated cells (p<0.05) and by 1.8±0.7 fold in the phenylephrine-treated cells (p<0.05) (Fig. 5).


Redox proteomics identification of oxidatively modified myocardial proteins in human heart failure: implications for protein function.

Brioschi M, Polvani G, Fratto P, Parolari A, Agostoni P, Tremoli E, Banfi C - PLoS ONE (2012)

Detection of protein carbonyls after immunoprecipitation of M-CK from cell lysate.HL-1 cells were treated with angiotensin II (AngII) or phenylephrine (PE) for 24 h, and the immunoprecipitates were immunoblotted with anti-DNP and anti-CK. Representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351458&req=5

pone-0035841-g005: Detection of protein carbonyls after immunoprecipitation of M-CK from cell lysate.HL-1 cells were treated with angiotensin II (AngII) or phenylephrine (PE) for 24 h, and the immunoprecipitates were immunoblotted with anti-DNP and anti-CK. Representative of three independent experiments.
Mentions: In comparison with the controls, M-CK protein carbonylation was significantly increased by 2.5±0.9 fold in the angiotensin II-treated cells (p<0.05) and by 1.8±0.7 fold in the phenylephrine-treated cells (p<0.05) (Fig. 5).

Bottom Line: The levels of carbonylated proteins were significantly higher in the HF patients than in the controls (p<0.01).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.

View Article: PubMed Central - PubMed

Affiliation: Centro Cardiologico Monzino IRCCS, Milan, Italy.

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.

Show MeSH
Related in: MedlinePlus