A 3.5-nm Structure of Rat TRPV4 Cation Channel Revealed by Zernike Phase-contrast Cryoelectron Microscopy

作者:Shigematsu Hideki; Sokabe Takaaki; Danev Radostin; Tominaga Makoto; Nagayama Kuniaki*
来源:JOURNAL OF BIOLOGICAL CHEMISTRY, 2010, 285(15): 11210-11218.
DOI:10.1074/jbc.M109.090712

摘要

The transient receptor potential vanilloid 4 (TRPV4) is a nonselective 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 phasecontrast 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.

  • 出版日期2010-4-9