JoVE Journal

Biochemistry

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Nanyang Technological University

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

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.

Hoofdstukken in deze video

0:04

Title

0:54

TRPV1 Mutagenesis

2:19

Generating the Recombinant Bacmid and Baculovirus for Protein Expression

5:14

eTRPV1 and Single-cysteine Mutant Purification

7:18

eTRPV1 Single-cysteine Mutant Site-directed Spin Labeling

8:12

eTRPV1 Spin-labeled Single Cysteine Mutant Reconstitution

9:30

Results: Characterization of the Minimal Cysteine-less TRPV1 Construct and Single-cysteine Mutants used for Spectroscopic Analysis

11:08

Conclusion

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