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PMID: 20044482 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A 3.5-nm structure of rat TRPV4 cation channel revealed by Zernike phase-contrast cryoelectron microscopy.

The Journal of biological chemistry ·Vol. 285 ·No. 15 ·2010-04-09 ·Pages 11210-8

Shigematsu H, Sokabe T, Danev R, Tominaga M, Nagayama K

Abstract

The transient receptor potential vanilloid 4 (TRPV4) is a non-selective cation channel responsive to various stimuli including cell swelling, warm temperatures (27-35 degrees C), and chemical compounds such as phorbol ester derivatives. Here we report the three-dimensional structure of full-length rat TRPV4 purified from baculovirus-infected Sf9 cells. Hexahistidine-tagged rat TRPV4 (His-rTRPV4) was solubilized with detergent and purified through affinity chromatography and size-exclusion chromatography. Chemical cross-linking analysis revealed that detergent-solubilized His-rTRPV4 was a tetramer. The 3.5-nm structure of rat TRPV4 was determined by cryoelectron microscopy using single-particle reconstruction from Zernike phase-contrast images. The overall structure comprises two distinct regions; a larger dense component, likely corresponding to the cytoplasmic N- and C-terminal regions, and a smaller component corresponding to the transmembrane region.

MeSH Terms
Animals Calcium/chemistry Cell Line Chromatography, Affinity/methods Cross-Linking Reagents/chemistry Cryoelectron Microscopy/methods Detergents/pharmacology Image Processing, Computer-Assisted Insecta Microscopy, Electron, Transmission/methods Microscopy, Phase-Contrast/methods Molecular Conformation Protein Conformation Rats TRPV Cation Channels/chemistry
Chemicals
Cross-Linking Reagents Detergents TRPV Cation Channels Trpv4 protein, rat Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Shigematsu Hideki
Division of Nano-Structure Physiology, Okazaki Institute for Integrative Bioscience, Higashiyama, Myodaiji, Okazaki 444-8787 Japan.
Sokabe Takaaki
Danev Radostin
Tominaga Makoto
Nagayama Kuniaki
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.090712
Published
2010-04-09
Epub
2009-00-31
Pages
11210-8
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2856998
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M109.090712 PMC Full Text View journal details
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