title: Identification of significant amino acids in multiple transmembrane domains of human transient receptor potential ankyrin 1 (TRPA1) for activation by eudesmol, an oxygenized sesquiterpene in hop essential oil.
abstract: Transient receptor potential ankyrin 1 (TRPA1) is a calcium-permeable non-selective cation channel that is activated by various noxious or irritant substances in nature, including spicy compounds. Many TRPA1 chemical activators have been reported; however, only limited information is available regarding the amino acid residues that contribute to the activation by non-electrophilic activators, whereas activation mechanisms by electrophilic ligands have been well characterized. We used intracellular Ca(2+) measurements and whole-cell patch clamp recordings to show that eudesmol, an oxygenated sesquiterpene present at high concentrations in the essential oil of hop cultivar Hallertau Hersbrucker, could activate human TRPA1. Gradual activation of inward currents with outward rectification by eudesmol was observed in human embryonic kidney-derived 293 cells expressing human TRPA1. This activation was completely blocked by a TRPA1-specific inhibitor, HC03-0031. We identified three critical amino acid residues in human TRPA1 in putative transmembrane domains 3, 4, and 5, namely threonine at 813, tyrosine at 840, and serine at 873, for activation by β-eudesmol in a systematic mutational study. Our results revealed a new TRPA1 activator in hop essential oil and provide a novel insight into mechanisms of human TRPA1 activation by non-electrophilic chemicals.
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and Kazuaki_Ohara@kirin.co.jp.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Ohara
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Fukuda
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Okada
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Sayoko
authors.initials: S
authors.lastname: Kitao
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ishida
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Kato
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Chika
authors.initials: C
authors.lastname: Takahashi
authors.affiliation: From the Research Laboratories for Health Science and Food Technologies, Kirin Company, Ltd., Yokohama, Kanagawa, 236-0004, Japan and.
authors.forename: Mikio
authors.initials: M
authors.lastname: Katayama
authors.affiliation: Division of Cell Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institute of Natural Sciences, Okazaki, Aichi, 444-8787, Japan.
authors.forename: Kunitoshi
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Division of Cell Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institute of Natural Sciences, Okazaki, Aichi, 444-8787, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Tominaga
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism*; D015220:Calcium Channels / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D027582:Humulus / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D009822:Oils, Volatile / Q000737:chemistry*; D012717:Sesquiterpenes / Q000737:chemistry*; D045787:Sesquiterpenes, Eudesmane / Q000737:chemistry* / Q000494:pharmacology*; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D015220:Calcium Channels; D009419:Nerve Tissue Proteins; D009822:Oils, Volatile; D012717:Sesquiterpenes; D045787:Sesquiterpenes, Eudesmane; D000074025:TRPA1 Cation Channel; C473350:TRPA1 protein, human; D050051:Transient Receptor Potential Channels; C051082:beta-eudesmol
keywords: Calcium Channel; Mutagenesis; Natural Product; Terpenoid; Transient Receptor Potential Channels (TRP Channels)