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Cardiac myosin binding protein C phosphorylation in cardiac disease.

Kuster DW, Bawazeer AC, Zaremba R, Goebel M, Boontje NM, van der Velden J - J. Muscle Res. Cell. Motil. (2011)

Bottom Line: Perturbations in sarcomeric function may in part underlie systolic and diastolic dysfunction of the failing heart.Sarcomeric dysfunction has been ascribed to changes in phosphorylation status of sarcomeric proteins caused by an altered balance between intracellular kinases and phosphatases during the development of cardiac disease.In the present review we discuss changes in phosphorylation of the thick filament protein myosin binding protein C (cMyBP-C) reported in failing myocardium, with emphasis on phosphorylation changes observed in familial hypertrophic cardiomyopathy caused by mutations in MYBPC3.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, VU University Medical Center, Amsterdam, The Netherlands. d.kuster@vumc.nl

ABSTRACT
Perturbations in sarcomeric function may in part underlie systolic and diastolic dysfunction of the failing heart. Sarcomeric dysfunction has been ascribed to changes in phosphorylation status of sarcomeric proteins caused by an altered balance between intracellular kinases and phosphatases during the development of cardiac disease. In the present review we discuss changes in phosphorylation of the thick filament protein myosin binding protein C (cMyBP-C) reported in failing myocardium, with emphasis on phosphorylation changes observed in familial hypertrophic cardiomyopathy caused by mutations in MYBPC3. Moreover, we will discuss assays which allow to distinguish between functional consequences of mutant sarcomeric proteins and (mal)adaptive changes in sarcomeric protein phosphorylation.

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Phosphorylation of cTnI and cMyBP-C by protein kinase C. Phosphorylation of cMyBP-C and cTnI after incubation of failing tissue samples with PKCα (n = 5) or PKCε (n = 2). Phosphorylation was assessed by ProQ Diamond stained gels and normalized to total SYPRO-stained cMyBP-C. Both PKC isoforms increased phosphorylation of cMyBP-C and cTnI, albeit with different specificities. *P < 0.05 versus baseline. BL baseline. Figure adapted from (Kooij et al. 2010a) with permission
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Fig4: Phosphorylation of cTnI and cMyBP-C by protein kinase C. Phosphorylation of cMyBP-C and cTnI after incubation of failing tissue samples with PKCα (n = 5) or PKCε (n = 2). Phosphorylation was assessed by ProQ Diamond stained gels and normalized to total SYPRO-stained cMyBP-C. Both PKC isoforms increased phosphorylation of cMyBP-C and cTnI, albeit with different specificities. *P < 0.05 versus baseline. BL baseline. Figure adapted from (Kooij et al. 2010a) with permission

Mentions: Protein kinase C can also phosphorylate cMyBP-C, as revealed by in vitro phosphorylation studies with recombinant PKC in vitro (Lim et al. 1985; Venema and Kuo 1993) or by PKC stimulation in intact cardiomyocytes (Venema and Kuo 1993). The phosphorylation sites of PKC on cMyBP-C overlap with those of PKA, as two of the three PKA-sites were also phosphorylated by PKC (Mohamed et al. 1998). PKC incubation combined with phosphorylation site-specific antibodies revealed that Ser273 and Ser302 are PKC target sites (Sadayappan et al. 2011). Protein kinase C is composed of a family of kinases, of which in the heart PKC-α is known to be upregulated in heart failure (Bowling et al. 1999). To study the effect of different PKC isoforms on cMyBP-C and cTnI phosphorylation, incubations with PKC-α and the novel–non Ca2+-activated-PKC-ε in skinned cardiomyocytes from failing tissue were performed (Kooij et al. 2010a). Both isoforms could phosphorylate cMyBP-C and cTnI, albeit with different substrate affinities (Fig. 4). Phosphorylation of cMyBP-C by another isoform of PKC, namely PKCζ, was shown by expressing a constitutively active form in cardiomyocytes (Wu and Solaro 2007). PKC phosphorylation of cMyBP-C is proposed to cause a decrease in actomyosin ATPase, which could be cardioprotective (Pyle et al. 2003).Fig. 4


Cardiac myosin binding protein C phosphorylation in cardiac disease.

Kuster DW, Bawazeer AC, Zaremba R, Goebel M, Boontje NM, van der Velden J - J. Muscle Res. Cell. Motil. (2011)

Phosphorylation of cTnI and cMyBP-C by protein kinase C. Phosphorylation of cMyBP-C and cTnI after incubation of failing tissue samples with PKCα (n = 5) or PKCε (n = 2). Phosphorylation was assessed by ProQ Diamond stained gels and normalized to total SYPRO-stained cMyBP-C. Both PKC isoforms increased phosphorylation of cMyBP-C and cTnI, albeit with different specificities. *P < 0.05 versus baseline. BL baseline. Figure adapted from (Kooij et al. 2010a) with permission
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Related In: Results  -  Collection

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Fig4: Phosphorylation of cTnI and cMyBP-C by protein kinase C. Phosphorylation of cMyBP-C and cTnI after incubation of failing tissue samples with PKCα (n = 5) or PKCε (n = 2). Phosphorylation was assessed by ProQ Diamond stained gels and normalized to total SYPRO-stained cMyBP-C. Both PKC isoforms increased phosphorylation of cMyBP-C and cTnI, albeit with different specificities. *P < 0.05 versus baseline. BL baseline. Figure adapted from (Kooij et al. 2010a) with permission
Mentions: Protein kinase C can also phosphorylate cMyBP-C, as revealed by in vitro phosphorylation studies with recombinant PKC in vitro (Lim et al. 1985; Venema and Kuo 1993) or by PKC stimulation in intact cardiomyocytes (Venema and Kuo 1993). The phosphorylation sites of PKC on cMyBP-C overlap with those of PKA, as two of the three PKA-sites were also phosphorylated by PKC (Mohamed et al. 1998). PKC incubation combined with phosphorylation site-specific antibodies revealed that Ser273 and Ser302 are PKC target sites (Sadayappan et al. 2011). Protein kinase C is composed of a family of kinases, of which in the heart PKC-α is known to be upregulated in heart failure (Bowling et al. 1999). To study the effect of different PKC isoforms on cMyBP-C and cTnI phosphorylation, incubations with PKC-α and the novel–non Ca2+-activated-PKC-ε in skinned cardiomyocytes from failing tissue were performed (Kooij et al. 2010a). Both isoforms could phosphorylate cMyBP-C and cTnI, albeit with different substrate affinities (Fig. 4). Phosphorylation of cMyBP-C by another isoform of PKC, namely PKCζ, was shown by expressing a constitutively active form in cardiomyocytes (Wu and Solaro 2007). PKC phosphorylation of cMyBP-C is proposed to cause a decrease in actomyosin ATPase, which could be cardioprotective (Pyle et al. 2003).Fig. 4

Bottom Line: Perturbations in sarcomeric function may in part underlie systolic and diastolic dysfunction of the failing heart.Sarcomeric dysfunction has been ascribed to changes in phosphorylation status of sarcomeric proteins caused by an altered balance between intracellular kinases and phosphatases during the development of cardiac disease.In the present review we discuss changes in phosphorylation of the thick filament protein myosin binding protein C (cMyBP-C) reported in failing myocardium, with emphasis on phosphorylation changes observed in familial hypertrophic cardiomyopathy caused by mutations in MYBPC3.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, VU University Medical Center, Amsterdam, The Netherlands. d.kuster@vumc.nl

ABSTRACT
Perturbations in sarcomeric function may in part underlie systolic and diastolic dysfunction of the failing heart. Sarcomeric dysfunction has been ascribed to changes in phosphorylation status of sarcomeric proteins caused by an altered balance between intracellular kinases and phosphatases during the development of cardiac disease. In the present review we discuss changes in phosphorylation of the thick filament protein myosin binding protein C (cMyBP-C) reported in failing myocardium, with emphasis on phosphorylation changes observed in familial hypertrophic cardiomyopathy caused by mutations in MYBPC3. Moreover, we will discuss assays which allow to distinguish between functional consequences of mutant sarcomeric proteins and (mal)adaptive changes in sarcomeric protein phosphorylation.

Show MeSH
Related in: MedlinePlus