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Related Experiment Videos

G protein function in the ischaemic myocardium.

L Will-Shahab1, W Rosenthal, W Schulze

  • 1Institute of Cardiovascular Research, Division of Cellular and Molecular Cardiology, Berlin-Buch, Germany.

European Heart Journal
|December 1, 1991
PubMed
Summary

Global ischemia impairs heart function by reducing specific G protein subtypes in sarcolemmal membranes. This study investigated G protein ADP-ribosylation in porcine hearts, revealing a decrease in PT-sensitive G proteins after 30 minutes of ischemia.

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Area of Science:

  • Cardiovascular Physiology
  • Molecular Cardiology
  • Biochemistry

Background:

  • Adenylyl cyclase (AC) activity, crucial for cyclic AMP production, is regulated by G proteins.
  • Ischemia impairs AC function in cardiac sarcolemmal preparations.
  • The role of G protein function in ischemia-induced AC impairment requires further investigation.

Purpose of the Study:

  • To investigate the contribution of G protein function to AC impairment during global ischemia.
  • To examine cholera toxin (CT)- and pertussis toxin (PT)-catalyzed ADP-ribosylation of G proteins in porcine heart sarcolemmal preparations subjected to global ischemia.

Main Methods:

  • Porcine hearts were subjected to global ischemia for varying durations (15-45 min).
  • Sarcolemmal preparations (SL) were isolated and analyzed for G protein ADP-ribosylation using CT and PT.

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  • Quantitative immunoblotting was employed to assess the levels of PT-sensitive G protein alpha-subunits.

    • Main Results:

    • CT-catalyzed ADP-ribosylation of a 45 kDa polypeptide showed no significant change after ischemia.
    • PT-catalyzed ADP-ribosylation of 39-41 kDa polypeptides significantly decreased after 30 min of ischemia (4.77 to 3.7 pmol mg-1 protein).
    • Quantitative immunoblotting indicated a loss of alpha-subunits of G0, Gi-1, and Gi-2 from sarcolemmal membranes, correlating with reduced PT-catalyzed ADP-ribosylation.

    Conclusions:

    • Global ischemia leads to a functional impairment of G proteins in cardiac sarcolemmal membranes.
    • The reduction in PT-sensitive G protein ADP-ribosylation is attributed to the loss of specific alpha-subunits (G0, Gi-1, Gi-2).
    • These G protein alterations likely contribute to the observed impairment of adenylyl cyclase function during ischemia.