The non-selective cation-permeable channel TRPC3 is a tetrahedron with a cap on the large cytoplasmic end

Biochem Biophys Res Commun. 2005 Aug 5;333(3):768-77. doi: 10.1016/j.bbrc.2005.05.181.

Abstract

TRPC3 plays important roles in neuronal differentiation and immune cell maturation by mediating the cationic current in response to phospholipase C activation, Ca2+ depletion, and diacylglycerol stimulation. Here, we purified the TRPC3 channel using a glycosylated tetramer and observed the structure using electron microscopy. Negatively stained specimens demonstrate homogeneous protein particles containing an internal cavity-like structure. These particle images were picked up by automated pick-up programs, aligned, and classified by the growing neural gas network method. Similarly oriented projections were averaged to decrease the signal-to-noise ratio. The averaged images progress from the top view to the side views, which are representative of their raw images. The top view confirmed the hypothesis of a four-domain structure, and the side view demonstrates a large cytoplasmic domain with a capped structure at the bottom, which is near a predicted locus of ion release. The total image of the protein is a blunt-edged trapezoid of 200 x 200 x 235 A. This large dimension of TRPC3 is also supported by the Stokes radius (92 A) obtained from gel filtration chromatography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Gel
  • Cytoplasm / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Ion Channels / chemistry
  • Ion Channels / metabolism*
  • Microscopy, Electron
  • Models, Molecular
  • TRPC Cation Channels

Substances

  • Ion Channels
  • TRPC Cation Channels
  • TRPC3 cation channel