title: Functional and Modeling Studies of the Transmembrane Region of the TRPM8 Channel.
abstract: Members of the transient receptor potential (TRP) ion channel family act as polymodal cellular sensors, which aid in regulating Ca(2+) homeostasis. Within the TRP family, TRPM8 is the cold receptor that forms a nonselective homotetrameric cation channel. In the absence of TRPM8 crystal structure, little is known about the relationship between structure and function. Inferences of TRPM8 structure have come from mutagenesis experiments coupled to electrophysiology, mainly regarding the fourth transmembrane helix (S4), which constitutes a moderate voltage-sensing domain, and about cold sensor and phosphatidylinositol 4,5-bisphosphate binding sites, which are both located in the C-terminus of TRPM8. In this study, we use a combination of molecular modeling and experimental techniques to examine the structure of the TRPM8 transmembrane and pore helix region including the conducting conformation of the selectivity filter. The model is consistent with a large amount of functional data and was further tested by mutagenesis. We present structural insight into the role of residues involved in intra- and intersubunit interactions and their link with the channel activity, sensitivity to icilin, menthol and cold, and impact on channel oligomerization.
authors.affiliation: Inserm, U1003, Laboratoire de Physiologie Cellulaire, Equipe labellisée par la Ligue contre le Cancer, Villeneuve d'Ascq, France; Laboratory of Excellence, Ion Channels Science and Therapeutics, Université de Lille 1, Villeneuve d'Ascq, France; Laboratoire Biophotonique Cellulaire Fonctionnelle. Institut de Recherche Interdisciplinaire, Villeneuve d'Ascq, France.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Bidaux
authors.affiliation: Centre for Cancer Research and Cell Biology, Queen's University of Belfast, Belfast, United Kingdom.
authors.forename: Miriam
authors.initials: M
authors.lastname: Sgobba
authors.affiliation: Inserm, U1003, Laboratoire de Physiologie Cellulaire, Equipe labellisée par la Ligue contre le Cancer, Villeneuve d'Ascq, France; Laboratory of Excellence, Ion Channels Science and Therapeutics, Université de Lille 1, Villeneuve d'Ascq, France.
authors.forename: Loic
authors.initials: L
authors.lastname: Lemonnier
authors.affiliation: Inserm, U1003, Laboratoire de Physiologie Cellulaire, Equipe labellisée par la Ligue contre le Cancer, Villeneuve d'Ascq, France; Laboratory of Excellence, Ion Channels Science and Therapeutics, Université de Lille 1, Villeneuve d'Ascq, France.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Borowiec
authors.affiliation: Inserm, U1003, Laboratoire de Physiologie Cellulaire, Equipe labellisée par la Ligue contre le Cancer, Villeneuve d'Ascq, France; Laboratory of Excellence, Ion Channels Science and Therapeutics, Université de Lille 1, Villeneuve d'Ascq, France.
authors.forename: Lucile
authors.initials: L
authors.lastname: Noyer
authors.affiliation: UCL School of Pharmacy, London, United Kingdom.
authors.forename: Srdan
authors.initials: S
authors.lastname: Jovanovic
authors.affiliation: Department of Biophysics, Educational and Scientific Centre, "Institute of Biology" Taras Shevchenko, Kiev National University, Kiev, Ukraine. Electronic address: a.zholos@althenia.org.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Zholos
authors.affiliation: UCL School of Pharmacy, London, United Kingdom. Electronic address: shozeb.haider@ucl.ac.uk.
authors.forename: Shozeb
authors.initials: S
authors.lastname: Haider
mesh_terms: D000595:Amino Acid Sequence; D019921:Biotinylation; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D018408:Patch-Clamp Techniques; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D050053:TRPM Cation Channels / Q000235:genetics / Q000378:metabolism*; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D050053:TRPM Cation Channels; C425966:TRPM8 protein, human; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human