Dissection of the components for PIP2 activation and thermosensation in TRP channels

Phosphatidylinositol 4,5-bisphosphate (PIP2) plays a central role in the activation of several transient receptor potential (TRP) channels. The role of PIP2 on temperature gating of thermoTRP channels has not been explored in detail, and the process of temperature activation is largely unexplained. In this work, we have exchanged different segments of the C-terminal region between cold-sensitive (TRPM8) and heat-sensitive (TRPV1) channels, trying to understand the role of the segment in PIP2 and temperature activation. A chimera in which the proximal part of the C-terminal of TRPV1 replaces an equivalent section of TRPM8 C-terminal is activated by PIP2 and confers the phenotype of heat activation. PIP2, but not temperature sensitivity, disappears when positively charged residues contained in the exchanged region are neutralized. Shortening the exchanged segment to a length of 11 aa produces voltage-dependent and temperature-insensitive channels. Our findings suggest the existence of different activation domains for temperature, PIP2, and voltage. We provide an interpretation for channel–PIP2 interaction using a full-atom molecular model of TRPV1 and PIP2 docking analysis.

[1]  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.

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

[3]  D. Hilgemann,et al.  The Complex and Intriguing Lives of PIP2 with Ion Channels and Transporters , 2001, Science's STKE.

[4]  G. Fernández-Ballester,et al.  Identification of a Tetramerization Domain in the C Terminus of the Vanilloid Receptor , 2004, The Journal of Neuroscience.

[5]  A. Basbaum,et al.  Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition , 2001, Nature.

[6]  D. Clapham,et al.  An introduction to TRP channels. , 2006, Annual review of physiology.

[7]  D. Julius,et al.  A Modular PIP2 Binding Site as a Determinant of Capsaicin Receptor Sensitivity , 2003, Science.

[8]  Beiying Liu,et al.  Functional Control of Cold- and Menthol-Sensitive TRPM8 Ion Channels by Phosphatidylinositol 4,5-Bisphosphate , 2005, The Journal of Neuroscience.

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

[10]  A. Stein,et al.  Phosphoinositide 3-Kinase Binds to TRPV1 and Mediates NGF-stimulated TRPV1 Trafficking to the Plasma Membrane , 2006, The Journal of general physiology.

[11]  T. Rohacs.  Regulation of TRP channels by PIP2 , 2007, Pflügers Archiv - European Journal of Physiology.

[12]  P. McNaughton,et al.  NGF rapidly increases membrane expression of TRPV1 heat‐gated ion channels , 2005, The EMBO journal.

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

[14]  B. Nilius,et al.  Regulation of TRP channels: a voltage-lipid connection. , 2007, Biochemical Society transactions.

[15]  E. Liman,et al.  Intracellular Ca2+ and the phospholipid PIP2 regulate the taste transduction ion channel TRPM5 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S. Cha,et al.  PIP2 Activates TRPV5 and Releases Its Inhibition by Intracellular Mg2+ , 2005, The Journal of general physiology.

[17]  I. Michailidis,et al.  PI(4,5)P2 regulates the activation and desensitization of TRPM8 channels through the TRP domain , 2005, Nature Neuroscience.

[18]  B. Nilius,et al.  Homo‐ and heterotetrameric architecture of the epithelial Ca2+ channels TRPV5 and TRPV6 , 2003, The EMBO journal.

[19]  M. Sansom,et al.  Molecular dynamics Simulation approaches to K channels: conformational flexibility and physiological function , 2005, IEEE Transactions on NanoBioscience.

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

[22]  D. Murray,et al.  Plasma membrane phosphoinositide organization by protein electrostatics , 2005, Nature.

[23]  L. Vyklický,et al.  Functional Role of C-Terminal Cytoplasmic Tail of Rat Vanilloid Receptor 1 , 2003, The Journal of Neuroscience.

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

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

[26]  Rich Olson,et al.  Structural basis for modulation and agonist specificity of HCN pacemaker channels , 2003, Nature.

[27]  D. Minor,et al.  The Pore Helix Dipole Has a Minor Role in Inward Rectifier Channel Function , 2005, Neuron.

[28]  B. Hille,et al.  Regulation of ion channels by phosphatidylinositol 4,5-bisphosphate , 2005, Current Opinion in Neurobiology.

[29]  G. Fernández-Ballester,et al.  Molecular architecture of the vanilloid receptor. Insights for drug design. , 2004, European journal of biochemistry.

[30]  D. Clapham,et al.  The TRPM7 channel is inactivated by PIP2 hydrolysis , 2002, Nature Cell Biology.

[31]  Haruhisa Okawa,et al.  Phosphatidylinositol 4,5-Bisphosphate Rescues TRPM4 Channels from Desensitization* , 2005, Journal of Biological Chemistry.