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Chapters
- 00:04Title
- 00:54TRPV1 Mutagenesis
- 02:19Generating the Recombinant Bacmid and Baculovirus for Protein Expression
- 05:14eTRPV1 and Single-cysteine Mutant Purification
- 07:18eTRPV1 Single-cysteine Mutant Site-directed Spin Labeling
- 08:12eTRPV1 Spin-labeled Single Cysteine Mutant Reconstitution
- 09:30Results: Characterization of the Minimal Cysteine-less TRPV1 Construct and Single-cysteine Mutants used for Spectroscopic Analysis
- 11:08Conclusion
This article describes specific methods to obtain biochemical quantities of detergent-solubilized TRPV1 for spectroscopic analysis. The combined protocols provide biochemical and biophysical tools that can be adapted to facilitate structural and functional studies for mammalian ion channels in a membrane-controlled environment.
