Temperature-induced opening of TRPV1 ion channel is stabilized by the pore domain

TRPV1 is the founding and best-studied member of the family of temperature-activated transient receptor potential ion channels (thermoTRPs). Voltage, chemicals and heat allosterically gate TRPV1. Molecular determinants of TRPV1 activation by capsaicin, allicin, acid, ammonia and voltage have been identified. However, the structures and mechanisms mediating TRPV1's pronounced temperature sensitivity remain unclear. Recent studies of the related channel TRPV3 identified residues in the pore region that are required for heat activation. We used both random and targeted mutagenesis screens of rat TRPV1 and identified point mutations in the outer pore region that specifically impair temperature activation. Single-channel analysis indicated that TRPV1 mutations disrupted heat sensitivity by ablating long channel openings, which are part of the temperature-gating pathway. We propose that sequential occupancy of short and long open states on activation provides a mechanism for enhancing temperature sensitivity. Our results suggest that the outer pore is important for the heat sensitivity of thermoTRPs.

[1]  H. Salazar,et al.  A single N-terminal cysteine in TRPV1 determines activation by pungent compounds from onion and garlic , 2008, Nature Neuroscience.

[2]  David Julius,et al.  Molecular Basis for Species-Specific Sensitivity to “Hot” Chili Peppers , 2002, Cell.

[3]  R. Latorre,et al.  Clues to understanding cold sensation: thermodynamics and electrophysiological analysis of the cold receptor TRPM8. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

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

[5]  L. Premkumar,et al.  Single‐channel properties of native and cloned rat vanilloid receptors , 2002, The Journal of physiology.

[6]  Ian M. Brooks,et al.  Extracellular Cations Sensitize and Gate Capsaicin Receptor TRPV1 Modulating Pain Signaling , 2005, The Journal of Neuroscience.

[7]  Jing Yao,et al.  Uncoupling Proton Activation of Vanilloid Receptor TRPV1 , 2007, The Journal of Neuroscience.

[8]  John B. Hogenesch,et al.  A Heat-Sensitive TRP Channel Expressed in Keratinocytes , 2002, Science.

[9]  Attila Toth,et al.  Molecular Determinants of Vanilloid Sensitivity in TRPV1* , 2004, Journal of Biological Chemistry.

[10]  Bernd Nilius,et al.  Gating of TRP channels: a voltage connection? , 2005, The Journal of physiology.

[11]  D. Julius,et al.  Acid potentiation of the capsaicin receptor determined by a key extracellular site. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Fan Yang,et al.  Thermosensitive TRP channel pore turret is part of the temperature activation pathway , 2010, Proceedings of the National Academy of Sciences.

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

[14]  David E. Clapham,et al.  TRPV3 is a calcium-permeable temperature-sensitive cation channel , 2002, Nature.

[15]  R. Latorre,et al.  Dissection of the components for PIP2 activation and thermosensation in TRP channels , 2007, Proceedings of the National Academy of Sciences.

[16]  S. Bingham,et al.  Vanilloid receptor-1 is essential for inflammatory thermal hyperalgesia , 2000, Nature.

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

[18]  A. Orth,et al.  High-throughput random mutagenesis screen reveals TRPM8 residues specifically required for activation by menthol , 2006, Nature Neuroscience.

[19]  F. Qin,et al.  Estimating single-channel kinetic parameters from idealized patch-clamp data containing missed events. , 1996, Biophysical journal.

[20]  R. Latorre,et al.  A Hot-Sensing Cold Receptor: C-Terminal Domain Determines Thermosensation in Transient Receptor Potential Channels , 2006, The Journal of Neuroscience.

[21]  M. Tominaga,et al.  TRPM2 activation by cyclic ADP‐ribose at body temperature is involved in insulin secretion , 2006, The EMBO journal.

[22]  Peter McIntyre,et al.  ANKTM1, a TRP-like Channel Expressed in Nociceptive Neurons, Is Activated by Cold Temperatures , 2003, Cell.

[23]  D. McKemy,et al.  Identification of a cold receptor reveals a general role for TRP channels in thermosensation , 2002, Nature.

[24]  A. Patapoutian,et al.  TRPV1 Is Activated by Both Acidic and Basic pH , 2009, The Journal of Neuroscience.

[25]  A. Auerbach,et al.  Maximum likelihood estimation of aggregated Markov processes , 1997, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[26]  Bernd Nilius,et al.  The principle of temperature-dependent gating in cold- and heat-sensitive TRP channels , 2004, Nature.

[27]  Manuela Schmidt,et al.  Pore region of TRPV3 ion channel is specifically required for heat activation , 2008, Nature Neuroscience.

[28]  Bernd Nilius,et al.  Heat activation of TRPM5 underlies thermal sensitivity of sweet taste , 2005, Nature.

[29]  B. Nilius,et al.  TRPM8 voltage sensor mutants reveal a mechanism for integrating thermal and chemical stimuli. , 2007, Nature chemical biology.

[30]  D. Kern,et al.  The role of dynamics in allosteric regulation. , 2003, Current opinion in structural biology.

[31]  Benjamin R. Myers,et al.  A Yeast Genetic Screen Reveals a Critical Role for the Pore Helix Domain in TRP Channel Gating , 2008, Neuron.

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

[33]  R. Latorre,et al.  ThermoTRP channels as modular proteins with allosteric gating. , 2007, Cell calcium.

[34]  José A. Matta,et al.  Voltage is a partial activator of rat thermosensitive TRP channels , 2007, The Journal of physiology.

[35]  Beiying Liu,et al.  Capsaicin activation of the pain receptor, VR1: multiple open states from both partial and full binding. , 2003, Biophysical journal.

[36]  B. Nilius,et al.  Neuronal TRP channels: thermometers, pathfinders and life-savers , 2008, Trends in Neurosciences.

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

[38]  P. Anand,et al.  TRPV3 is a temperature-sensitive vanilloid receptor-like protein , 2002, Nature.

[39]  P. McIntyre,et al.  A TRP Channel that Senses Cold Stimuli and Menthol , 2002, Cell.

[40]  B. Hille,et al.  Ionic channels of excitable membranes , 2001 .

[41]  Beiying Liu,et al.  Thermodynamics of heat activation of single capsaicin ion channels VR1. , 2003, Biophysical journal.

[42]  Makoto Tominaga,et al.  Heat-Evoked Activation of the Ion Channel, TRPV4 , 2002, The Journal of Neuroscience.