title: Polyester modification of the mammalian TRPM8 channel protein: implications for structure and function.
abstract: The TRPM8 ion channel is expressed in sensory neurons and is responsible for sensing environmental cues, such as cold temperatures and chemical compounds, including menthol and icilin. The channel functional activity is regulated by various physical and chemical factors and is likely to be preconditioned by its molecular composition. Our studies indicate that the TRPM8 channel forms a structural-functional complex with the polyester poly-(R)-3-hydroxybutyrate (PHB). We identified by mass spectrometry a number of PHB-modified peptides in the N terminus of the TRPM8 protein and in its extracellular S3-S4 linker. Removal of PHB by enzymatic hydrolysis and site-directed mutagenesis of both the serine residues that serve as covalent anchors for PHB and adjacent hydrophobic residues that interact with the methyl groups of the polymer resulted in significant inhibition of TRPM8 channel activity. We conclude that the TRPM8 channel undergoes posttranslational modification by PHB and that this modification is required for its normal function.
authors.affiliation: New Jersey Medical School, UMDNJ, Department of Pharmacology and Physiology, 185 South Orange Avenue, MSB H626, Newark NJ 07103, USA.
authors.forename: Chike
authors.initials: C
authors.lastname: Cao
authors.affiliation: New Jersey Medical School, UMDNJ, Department of Pharmacology and Physiology, 185 South Orange Avenue, MSB H626, Newark NJ 07103, USA.
authors.forename: Yevgen
authors.initials: Y
authors.lastname: Yudin
authors.affiliation: New Jersey Medical School, UMDNJ, Department of Pharmacology and Physiology, 185 South Orange Avenue, MSB H626, Newark NJ 07103, USA.
authors.forename: Yann
authors.initials: Y
authors.lastname: Bikard
authors.affiliation: New Jersey Medical School Cancer Center - UMDNJ, Center for Advanced Proteomics Research, Building F1105, 205 South Orange Avenue, Newark, NJ 07103 USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: New Jersey Medical School Cancer Center - UMDNJ, Center for Advanced Proteomics Research, Building F1105, 205 South Orange Avenue, Newark, NJ 07103 USA.
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.affiliation: New Jersey Medical School Cancer Center - UMDNJ, Center for Advanced Proteomics Research, Building F1105, 205 South Orange Avenue, Newark, NJ 07103 USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.affiliation: Universität Stuttgart Zentrum für Bioverfahrenstechnik Institut für Mikrobiologie, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Jendrossek
authors.affiliation: Dalhousie University, Proteomics Core Facility, Clinical Research Centre, Room C-304 5849 University Avenue, PO Box 15000, Halifax, NS, B3H 4R2 Canada.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Cohen
authors.affiliation: Dalhousie University, Department of Physiology and Biophysics Faculty of Medicine, Sir Charles Tupper Medical Building, Halifax, Room 5G, 5850 College St., Halifax, NS, B3H 4R2 Canada.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Pavlov
authors.affiliation: New Jersey Medical School, UMDNJ, Department of Pharmacology and Physiology, 185 South Orange Avenue, MSB H626, Newark NJ 07103, USA.
authors.forename: Tibor
authors.initials: T
authors.lastname: Rohacs
authors.affiliation: New Jersey Medical School, UMDNJ, Department of Pharmacology and Physiology, 185 South Orange Avenue, MSB H626, Newark NJ 07103, USA.|University of Illinois College of Medicine, Department of Cancer Biology and Pharmacology, 1 Illini Drive, Peoria, IL 61605, USA.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Zakharian
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D018929:Cell Culture Techniques; D057809:HEK293 Cells; D006801:Humans; D006885:Hydroxybutyrates / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011091:Polyesters / Q000378:metabolism*; D000091402:Prohibitins; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013329:Structure-Activity Relationship; D050053:TRPM Cation Channels / Q000235:genetics / Q000378:metabolism*; D000094122:Polyhydroxybutyrates
chemical_list: D006885:Hydroxybutyrates; D011091:Polyesters; D000091402:Prohibitins; D050053:TRPM Cation Channels; C000717328:PHB protein, human; C425966:TRPM8 protein, human; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates