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Effect of transmembrane Ca2+ gradient on Gs function

G F Fan1, Y G Huang, Y H Bai

  • 1National Laboratory of Biomacromolecules, Chinese Academy of Sciences, Beijing.

FEBS Letters
|January 2, 1995
PubMed
Summary
This summary is machine-generated.

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A physiological calcium (Ca2+) gradient across cell membranes enhances Gs protein activity and adenylate cyclase stimulation. This membrane fluidity modulation is key for optimal G protein function.

Area of Science:

  • Neurobiology
  • Membrane Biophysics
  • Signal Transduction

Background:

  • G protein-coupled receptors (GPCRs) and their downstream effectors, like adenylate cyclase, are crucial for cellular communication.
  • Transmembrane ion gradients, particularly for calcium (Ca2+), play a significant role in regulating cellular processes.
  • The physical state of the cell membrane, including its fluidity, can impact protein function.

Purpose of the Study:

  • To investigate the effect of a transmembrane Ca2+ gradient on the activity of Gs proteins and adenylate cyclase.
  • To explore the relationship between membrane physical state (fluidity) and Gs protein function in proteoliposomes.
  • To determine if physiological Ca2+ gradients are essential for optimal Gs-mediated signaling.

Main Methods:

Related Experiment Videos

  • Co-reconstitution of bovine brain Gs and adenylate cyclase into asolectin liposomes.
  • Creation of proteoliposomes with physiological and inverse transmembrane Ca2+ gradients.
  • Measurement of Gs activity (GTPγS binding and adenylate cyclase stimulation).
  • Assessment of membrane physical state using time-resolved fluorescence anisotropy of diphenylhexatriene (DPH).

    • Main Results:

    • Gs activity, including GTPγS binding and adenylate cyclase stimulation, was highest in proteoliposomes with a physiological Ca2+ gradient.
    • Activity was significantly lower when the Ca2+ gradient was in the inverse direction.
    • Dissipation of the Ca2+ gradient using A23187 diminished these differences.
    • A proper transmembrane Ca2+ gradient was correlated with increased membrane fluidity.

    Conclusions:

    • A physiological transmembrane Ca2+ gradient is essential for optimal Gs protein function, enhancing both GTP-binding and adenylate cyclase stimulation.
    • Increased membrane fluidity, promoted by a proper Ca2+ gradient, may be the mechanism underlying enhanced Gs activity.
    • These findings highlight the importance of calcium gradients in regulating G protein-mediated signaling pathways.