Production and purification of TRPV2 and TRPV5 for structural and functional studies
| Autor: | Edwin C. Fluck, Ruth A. Pumroy, Vera Y. Moiseenkova-Bell |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0303 health sciences
TRPV5 biology Chemistry 030303 biophysics Saccharomyces cerevisiae TRPV2 TRPV Cation Channels biology.organism_classification Article Epitope Rats 03 medical and health sciences Transient receptor potential channel Affinity chromatography Biochemistry biology.protein Animals Rabbits Heterologous expression Antibody |
| Zdroj: | Methods in Enzymology ISBN: 9780323853767 Methods Enzymol |
| DOI: | 10.1016/bs.mie.2021.02.007 |
| Popis: | The transient receptor potential (TRP) vanilloid 2 (TRPV2) and TRP vanilloid 5 (TRPV5) cation channels play an important role in various physiological and pathophysiological processes. The heterologous expression and purification of these channels is critical for functional and structural characterization of these important proteins. Full-length rat TRPV2 and rabbit TRPV5 can both be expressed in Saccharomyces cerevisiae and affinity purified using the 1D4 epitope and antibody to yield pure, functional channels. Further, these channels can be reconstituted into lipid nanodiscs for a more functionally relevant environment. Presented here are protocols for the expression of full-length rat TRPV2 and rabbit TRPV5 in Saccharomyces cerevisiae, their affinity purification, and their reconstitution into nanodiscs for structural and functional studies. |
| Databáze: | OpenAIRE |
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