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Related Experiment Videos

Protein phosphorylation and cardiac function: cholinergic-adrenergic interaction

S Bartel1, P Karczewski, E G Krause

  • 1Max Delbrück Centre for Molecular Medicine, Molecular Cardiology, Berlin-Buch, Germany.

Cardiovascular Research
|November 1, 1993
PubMed
Summary

Cholinergic agonists like carbachol reduce cardiac protein phosphorylation, independent of cyclic AMP (cAMP) levels. This suggests a role for cyclic guanosine monophosphate (cGMP) in carbachol's anti-adrenergic effects in the heart.

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Area of Science:

  • Cardiology
  • Molecular Pharmacology
  • Biochemistry

Background:

  • Muscarinic inhibition of cardiac contractility may involve mechanisms beyond cyclic AMP (cAMP) reduction, potentially affecting cAMP-linked protein phosphorylation.
  • Cholinergic agonists might antagonize agents that activate the cAMP signaling pathway at the protein phosphorylation level.

Purpose of the Study:

  • To investigate the effect of carbachol on in vivo phosphorylation of key cardiac proteins (phospholamban, troponin I, 15 kDa protein).
  • To determine if carbachol antagonizes cAMP-mediated protein phosphorylation during positive inotropic interventions.

Main Methods:

  • Isolated rat hearts were used to monitor contractile activity during drug administration.
  • Hearts were freeze-clamped for analysis of protein phosphorylation and metabolites, including cAMP and cyclic guanosine monophosphate (cGMP).

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  • A 'back phosphorylation' method was employed to quantify in vivo phosphorylation levels.

    • Main Results:

    • Carbachol attenuated the in vivo phosphorylation of all studied phosphoproteins induced by cAMP-elevating drugs.
    • cAMP levels were unaffected or decreased by isoprenaline, yet carbachol still reduced phosphorylation.
    • Carbachol reversed cAMP-mediated phosphorylase a activation, and sodium nitroprusside also reduced phosphorylation and contractile force.

    Conclusions:

    • Carbachol reduces cAMP-mediated cardiac protein phosphorylation, acting on targets downstream of cAMP generation.
    • The anti-adrenergic action of carbachol in the myocardium appears to involve cyclic guanosine monophosphate (cGMP).
    • Findings highlight the complex interplay between cAMP, cGMP, and protein phosphorylation in regulating cardiac function.