Related Experiment Video

Updated: Feb 5, 2026

Optimized Transfection Strategy for Expression and Electrophysiological Recording of Recombinant Voltage-Gated Ion Channels in HEK-293T Cells
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Optimized Transfection Strategy for Expression and Electrophysiological Recording of Recombinant Voltage-Gated Ion Channels in HEK-293T Cells

Published on: January 19, 2011

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In vitro and in vivo phosphorylation of the Cav2.3 voltage-gated R-type calcium channel

T Schneider1, S Alpdogan, J Hescheler

  • 1a Center of Physiology and Pathophysiology , Institute of Neurophysiology , Cologne , Germany.

Channels (Austin, Tex.)
|September 1, 2018
PubMed

Insights

Adenosine triphosphate (ATP) depletion causes Cav2.3 channel rundown, but ATP can prevent this. Protein phosphorylation, regulated by ATP, is crucial for Cav2.3 channel function and gating.

Area of Science:

  • Molecular biology
  • Neuroscience
  • Ion channel physiology

Background:

  • HEK-293 cells express recombinant human Cav2.3+β3 channel subunits.
  • Whole-cell current recordings reveal channel behavior.
  • Adenosine triphosphate (ATP) depletion affects channel function.

Purpose of the Study:

  • To investigate the role of ATP in Cav2.3 channel function.
  • To understand the mechanism of channel rundown.
  • To explore the impact of protein phosphorylation on Cav2.3 channels.

Main Methods:

  • Whole-cell patch-clamp recordings in HEK-293 cells.
  • Application of hydrolyzable ATP to counteract channel rundown.
  • Inhibition of serine/threonine, tyrosine, and lipid kinases.

Main Results:

  • ATP depletion leads to Cav2.3 channel rundown and a shift in voltage-dependent gating.
  • Hydrolyzable ATP prevents channel rundown.
  • Inhibition of serine/threonine kinases abolishes ATP's protective effect.
  • Protein phosphorylation is essential for Cav2.3 channel function.

Conclusions:

  • ATP-dependent protein phosphorylation regulates Cav2.3 channel function and gating.
  • Channel rundown is linked to dephosphorylation during cell dialysis.
  • Phosphorylation is critical for maintaining normal Cav2.3 channel activity.
Keywords:
Covalent modificationexon skippingfacilitationprotein kinase Aprotein kinase Csplice variants

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