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1Department of Physiology,David Geffen School of Medicine, University of California, Los Angeles, 2Department of Anesthesiology,David Geffen School of Medicine, University of California, Los Angeles, 3Department of Anesthesiology, Medicine and Physiology,David Geffen School of Medicine, University of California, Los Angeles
Chapters
- 00:00Title
- 01:01Introduction
- 01:35Subcloning, Expressing, and Binding a GST-Fusion protein to GLU-coated beads
- 06:41The GST-fusion Protein Binding Assay
- 09:12Preparing gel-separated proteins for Mass Spectrometry
- 15:33Representative Results/ Outcome
- 16:03Conclusion
We describe a simple protocol to identify brain proteins that bind to the full length C terminus of ATP-gated P2X2 receptors. The extension and systematic application of this approach to all P2X receptors is expected to lead to a better understanding of P2X receptor signaling.
Tags
ProteomicsProteinsP2X2 Ligand-gated Cation ChannelsLigand-gated Ion ChannelsSynaptic CommunicationNervous SystemCys LoopGlutamate-gated ChannelsP2X Receptor ChannelsTransmitter BindingPore OpeningIons FlowElectrochemical GradientsCalcium IonsDownstream Signaling RolesGene RegulationProtein Signaling ComplexesC-terminal AspectsP2X2 Receptor Cytosolic DomainATP-gated Cation ChannelsSubunitsExcitatory Synaptic TransmissionNeurotransmitter ReleaseExcitable CellsNon-excitable CellsNeuronal SignalingInflammationCardiovascular Function
