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Human pannexin 1 channel is not phosphorylated by Src tyrosine kinase at Tyr199 and Tyr309

Zheng Ruan1, Junuk Lee1, Yangyang Li1

  • 1Department of Structural Biology, Van Andel Institute, Grand Rapids, United States.

Elife
|May 23, 2024
PubMed

Insights

Commercially available antibodies for Pannexin 1 (PANX1) phosphorylation are nonspecific. Re-examination of the tyrosine phosphorylation-dependent activation of the PANX1 channel is needed.

Area of Science:

  • Molecular biology
  • Cellular signaling
  • Ion channel function

Background:

  • Protein phosphorylation regulates protein activity and function.
  • Pannexin 1 (PANX1) channels are implicated in cell death, inflammation, and muscle contraction.
  • Src kinase-mediated PANX1 tyrosine phosphorylation is proposed to activate the channel.

Purpose of the Study:

  • To validate the role of specific tyrosine phosphorylation sites in PANX1 channel activation.
  • To assess the specificity of antibodies used to detect PANX1 phosphorylation.
  • To re-evaluate the current model of Src-dependent PANX1 channel activation.

Main Methods:

  • Testing the specificity of commercial antibodies against phosphorylated PANX1 Tyr198/Tyr308 (mouse).
  • Investigating Src kinase phosphorylation of PANX1 in heterologous expression systems.
  • Analyzing endogenous PANX1 phosphorylation using validated methods.

Main Results:

  • Commercial antibodies targeting PANX1 phosphorylation sites Tyr198/Tyr308 are nonspecific.
  • Neither Tyr198 nor Tyr308 is a major phosphorylation site for Src kinase in tested systems.
  • Existing data on PANX1 phosphorylation may require reinterpretation.

Conclusions:

  • The specificity of commonly used antibodies for PANX1 phosphorylation is questionable.
  • The proposed mechanism of Src-dependent PANX1 activation via Tyr198/308 phosphorylation needs reassessment.
  • Further research is required to elucidate PANX1 channel regulation.
Keywords:
Pannexin 1Src kinaseY199Y309biochemistrychemical biologyhumanmousetyrosine phosphorylation

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