Related Experiment Videos
Proteomic and functional evidence for a P2X7 receptor signalling complex.
1Institute of Molecular Physiology, University of Sheffield, Alfred Denny Building, Western Bank, Sheffield S10 2TN, UK.
The EMBO Journal
|November 15, 2001
Summary
Researchers identified proteins interacting with the P2X7 receptor, crucial for ATP-gated ion channels. This interaction influences cytoskeletal changes and receptor activity via phosphorylation and dephosphorylation.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- P2X receptors are ATP-gated ion channels.
- P2X7 receptor activation triggers cytoskeletal rearrangements like membrane blebbing.
Purpose of the Study:
- Identify proteins interacting with the P2X7 receptor.
- Investigate the role of these interactions in P2X7 receptor function and downstream signaling.
Main Methods:
- Affinity purification coupled with mass spectrometry and immunoblotting.
- Site-directed mutagenesis to identify phosphorylation sites.
- Whole-cell electrophysiology recordings.
Main Results:
- Identified 11 interacting proteins, including RPTPbeta, beta-actin, and integrin beta2.
- P2X7 receptor activation led to its dephosphorylation at Tyr(343).
- Dephosphorylation reduced ionic currents and slowed membrane blebbing.
Conclusions:
- A P2X7 receptor signaling complex was identified.
- Interacting proteins may mediate cytoskeletal rearrangements.
- RPTPbeta may provide feedback control through dephosphorylation.