title: Implications of Human Transient Receptor Potential Melastatin 8 (TRPM8) Channel Gating from Menthol Binding Studies of the Sensing Domain.
abstract: The transient receptor potential melastatin 8 (TRPM8) ion channel is the primary cold sensor in humans. TRPM8 is gated by physiologically relevant cold temperatures and chemical ligands that induce cold sensations, such as the analgesic compound menthol. Characterization of TRPM8 ligand-gated channel activation will lead to a better understanding of the fundamental mechanisms that underlie TRPM8 function. Here, the direct binding of menthol to the isolated hTRPM8 sensing domain (transmembrane helices S1-S4) is investigated. These data are compared with two mutant sensing domain proteins, Y745H (S2 helix) and R842H (S4 helix), which have been previously identified in full length TRPM8 to be menthol insensitive. The data presented herein show that menthol specifically binds to the wild type, Y745H, and R842H TRPM8 sensing domain proteins. These results are the first to show that menthol directly binds to the TRPM8 sensing domain and indicates that Y745 and R842 residues, previously identified in functional studies as crucial to menthol sensitivity, do not affect menthol binding but instead alter coupling between the sensing domain and the pore domain.
authors.affiliation: School of Molecular Sciences, Arizona State University , 551 E. University Drive, Tempe, Arizona 85287, United States.|The Biodesign Institute, Arizona State University , Tempe, Arizona 85281, United States.|The Virginia G. Piper Center for Personalized Diagnostics, Arizona State University , Tempe, Arizona 85281, United States.|The Magnetic Resonance Research Center, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Parthasarathi
authors.initials: P
authors.lastname: Rath
authors.affiliation: School of Molecular Sciences, Arizona State University , 551 E. University Drive, Tempe, Arizona 85287, United States.|The Biodesign Institute, Arizona State University , Tempe, Arizona 85281, United States.|The Virginia G. Piper Center for Personalized Diagnostics, Arizona State University , Tempe, Arizona 85281, United States.|The Magnetic Resonance Research Center, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Jacob K
authors.initials: JK
authors.lastname: Hilton
authors.affiliation: School of Molecular Sciences, Arizona State University , 551 E. University Drive, Tempe, Arizona 85287, United States.|The Biodesign Institute, Arizona State University , Tempe, Arizona 85281, United States.|The Virginia G. Piper Center for Personalized Diagnostics, Arizona State University , Tempe, Arizona 85281, United States.|The Magnetic Resonance Research Center, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Sisco
authors.affiliation: School of Molecular Sciences, Arizona State University , 551 E. University Drive, Tempe, Arizona 85287, United States.|The Biodesign Institute, Arizona State University , Tempe, Arizona 85281, United States.|The Virginia G. Piper Center for Personalized Diagnostics, Arizona State University , Tempe, Arizona 85281, United States.|The Magnetic Resonance Research Center, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Wade D
authors.initials: WD
authors.lastname: Van Horn
mesh_terms: D001665:Binding Sites; D057809:HEK293 Cells; D006801:Humans; D008610:Menthol / Q000378:metabolism*; D008958:Models, Molecular; D017354:Point Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D050053:TRPM Cation Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D050053:TRPM Cation Channels; C425966:TRPM8 protein, human; D008610:Menthol