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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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Immunofluorescence analysis of carbonylated proteins in cardiomyocytes after treatment with angiotensin II (AngII) or phenylephrine (PE).The cells were stained with anti-DNP antibody and visualised by means of a secondary antibody conjugated with Alexa Fluor dye 488. Representative of three independent experiments.
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pone-0035841-g004: Immunofluorescence analysis of carbonylated proteins in cardiomyocytes after treatment with angiotensin II (AngII) or phenylephrine (PE).The cells were stained with anti-DNP antibody and visualised by means of a secondary antibody conjugated with Alexa Fluor dye 488. Representative of three independent experiments.

Mentions: We then looked for the presence of protein carbonyls in HL-1 cells exposed for different times to angiotensin II or phenylephrine, the most potent H2O2 inductors under our experimental conditions. As shown in Figure 4, carbonyls were clearly detectable in the HL-1 cells incubated with both agents after 1 h, and remained detectable for up to 24 h.


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)

Immunofluorescence analysis of carbonylated proteins in cardiomyocytes after treatment with angiotensin II (AngII) or phenylephrine (PE).The cells were stained with anti-DNP antibody and visualised by means of a secondary antibody conjugated with Alexa Fluor dye 488. Representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0035841-g004: Immunofluorescence analysis of carbonylated proteins in cardiomyocytes after treatment with angiotensin II (AngII) or phenylephrine (PE).The cells were stained with anti-DNP antibody and visualised by means of a secondary antibody conjugated with Alexa Fluor dye 488. Representative of three independent experiments.
Mentions: We then looked for the presence of protein carbonyls in HL-1 cells exposed for different times to angiotensin II or phenylephrine, the most potent H2O2 inductors under our experimental conditions. As shown in Figure 4, carbonyls were clearly detectable in the HL-1 cells incubated with both agents after 1 h, and remained detectable for up to 24 h.

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