Structure of TRPV1 channel revealed by electron cryomicroscopy

The transient receptor potential (TRP) family of ion channels participate in many signaling pathways. TRPV1 functions as a molecular integrator of noxious stimuli, including heat, low pH, and chemical ligands. Here, we report the 3D structure of full-length rat TRPV1 channel expressed in the yeast Saccharomyces cerevisiae and purified by immunoaffinity chromatography. We demonstrate that the recombinant purified TRPV1 channel retains its structural and functional integrity and is suitable for structural analysis. The 19-Å structure of TRPV1 determined by using single-particle electron cryomicroscopy exhibits fourfold symmetry and comprises two distinct regions: a large open basket-like domain, likely corresponding to the cytoplasmic N- and C-terminal portions, and a more compact domain, corresponding to the transmembrane portion. The assignment of transmembrane and cytoplasmic regions was supported by fitting crystal structures of the structurally homologous Kv1.2 channel and isolated TRPV1 ankyrin repeats into the TRPV1 structure.

[1]  Y. Kubo,et al.  The Motor Protein Prestin Is a Bullet-shaped Molecule with Inner Cavities* , 2008, Journal of Biological Chemistry.

[2]  Y. Mori,et al.  Three-dimensional Reconstruction Using Transmission Electron Microscopy Reveals a Swollen, Bell-shaped Structure of Transient Receptor Potential Melastatin Type 2 Cation Channel* , 2007, Journal of Biological Chemistry.

[3]  Matthew D. Pagel,et al.  The structure of the prokaryotic cyclic nucleotide-modulated potassium channel MloK1 at 16 A resolution. , 2007, Structure.

[4]  Xiangshu Jin,et al.  The Ankyrin Repeats of TRPV1 Bind Multiple Ligands and Modulate Channel Sensitivity , 2007, Neuron.

[5]  Y. Hiroaki,et al.  The TRPC3 channel has a large internal chamber surrounded by signal sensing antennas. , 2007, Journal of molecular biology.

[6]  Andreas Kreusch,et al.  Crystal structure of the human TRPV2 channel ankyrin repeat domain , 2006, Protein science : a publication of the Protein Society.

[7]  Rachelle Gaudet,et al.  Structure of the N-terminal Ankyrin Repeat Domain of the TRPV2 Ion Channel* , 2006, Journal of Biological Chemistry.

[8]  A. Patapoutian,et al.  Trp ion channels and temperature sensation. , 2006, Annual review of neuroscience.

[9]  Bernd Nilius,et al.  Permeation and selectivity of TRP channels. , 2006, Annual review of physiology.

[10]  Y. Hara,et al.  The non-selective cation-permeable channel TRPC3 is a tetrahedron with a cap on the large cytoplasmic end. , 2005, Biochemical and biophysical research communications.

[11]  E. Campbell,et al.  Crystal Structure of a Mammalian Voltage-Dependent Shaker Family K+ Channel , 2005, Science.

[12]  Wah Chiu,et al.  The pore structure of the closed RyR1 channel. , 2005, Structure.

[13]  Bernd Nilius,et al.  TRP channels: a TR(I)P through a world of multifunctional cation channels , 2005, Pflügers Archiv.

[14]  B. Nilius,et al.  Sensing with TRP channels , 2005, Nature chemical biology.

[15]  B. Niemeyer,et al.  Structure-function analysis of TRPV channels , 2005, Naunyn-Schmiedeberg's Archives of Pharmacology.

[16]  Craig Montell,et al.  The TRP Superfamily of Cation Channels , 2005, Science's STKE.

[17]  O. Sokolova.  Structure of cation channels, revealed by single particle electron microscopy , 2004, FEBS letters.

[18]  David E. Clapham,et al.  TRP channels as cellular sensors , 2003, Nature.

[19]  N. Grigorieff,et al.  Conformational changes in the C terminus of Shaker K+ channel bound to the rat Kvβ2-subunit , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  H. Hellmich,et al.  Overexpression and purification of the vanilloid receptor in yeast (Saccharomyces cerevisiae). , 2003, Biochemical and biophysical research communications.

[21]  Wah Chiu,et al.  Structure of the Type 1 Inositol 1,4,5-Trisphosphate Receptor Revealed by Electron Cryomicroscopy* , 2003, Journal of Biological Chemistry.

[22]  M. Cadene,et al.  X-ray structure of a voltage-dependent K+ channel , 2003, Nature.

[23]  Youxing Jiang,et al.  Crystal structure and mechanism of a calcium-gated potassium channel , 2002, Nature.

[24]  G. Schertler,et al.  Molecular architecture of a retinal cGMP‐gated channel: the arrangement of the cytoplasmic domains , 2002, The EMBO journal.

[25]  B. Minke,et al.  TRP channel proteins and signal transduction. , 2002, Physiological reviews.

[26]  R. Dutzler,et al.  X-ray structure of a ClC chloride channel at 3.0 Å reveals the molecular basis of anion selectivity , 2002, Nature.

[27]  Peter M. Blumberg,et al.  Analysis of the Native Quaternary Structure of Vanilloid Receptor 1* 210 , 2001, The Journal of Biological Chemistry.

[28]  N. Grigorieff,et al.  Three-dimensional structure of a voltage-gated potassium channel at 2.5 nm resolution. , 2001, Structure.

[29]  D. Julius,et al.  The vanilloid receptor: a molecular gateway to the pain pathway. , 2001, Annual review of neuroscience.

[30]  D. Julius,et al.  Capsaicin receptor in the pain pathway. , 2000, Japanese journal of pharmacology.

[31]  A I Basbaum,et al.  Impaired nociception and pain sensation in mice lacking the capsaicin receptor. , 2000, Science.

[32]  R. Nakamoto,et al.  Use of chemical chaperones in the yeast Saccharomyces cerevisiae to enhance heterologous membrane protein expression: high-yield expression and purification of human P-glycoprotein. , 2000, Archives of biochemistry and biophysics.

[33]  W Chiu,et al.  EMAN: semiautomated software for high-resolution single-particle reconstructions. , 1999, Journal of structural biology.

[34]  D. Oprian,et al.  Tertiary interactions between transmembrane segments 3 and 5 near the cytoplasmic side of rhodopsin. , 1999, Biochemistry.

[35]  A. Basbaum,et al.  The Cloned Capsaicin Receptor Integrates Multiple Pain-Producing Stimuli , 1998, Neuron.

[36]  B. Chait,et al.  The structure of the potassium channel: molecular basis of K+ conduction and selectivity. , 1998, Science.

[37]  D. Julius,et al.  The capsaicin receptor: a heat-activated ion channel in the pain pathway , 1997, Nature.

[38]  L. Stryer,et al.  Kinetics of calcium channel opening by inositol 1,4,5-trisphosphate. , 1990, Biochemistry.

[39]  G. Rubin,et al.  Molecular characterization of the drosophila trp locus: A putative integral membrane protein required for phototransduction , 1989, Neuron.

[40]  J. Dubochet,et al.  Cryo-electron microscopy of vitrified specimens , 1988, Quarterly Reviews of Biophysics.

[41]  P. Hargrave,et al.  Localization of binding sites for carboxyl terminal specific anti-rhodopsin monoclonal antibodies using synthetic peptides. , 1984, Biochemistry.

[42]  R. Molday,et al.  Monoclonal antibodies to rhodopsin: characterization, cross-reactivity, and application as structural probes. , 1983, Biochemistry.

[43]  A. Manning,et al.  Abnormal Electroretinogram from a Drosophila Mutant , 1969, Nature.