title: Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells.
abstract: The controlled activation of proteins in living cells is an important goal in protein-design research, but to introduce an artificial activation switch into membrane proteins through rational design is a significant challenge because of the structural and functional complexity of such proteins. Here we report the allosteric activation of two types of membrane-bound neurotransmitter receptors, the ion-channel type and the G-protein-coupled glutamate receptors, using coordination chemistry in living cells. The high programmability of coordination chemistry enabled two His mutations, which act as an artificial allosteric site, to be semirationally incorporated in the vicinity of the ligand-binding pockets. Binding of Pd(2,2'-bipyridine) at the allosteric site enabled the active conformations of the glutamate receptors to be stabilized. Using this approach, we were able to activate selectively a mutant glutamate receptor in live neurons, which initiated a subsequent signal-transduction pathway.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.|Department of Technology and Ecology, Hall of Global Environmental Studies, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kiyonaka
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
authors.forename: Ryou
authors.initials: R
authors.lastname: Kubota
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Michibata
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehirocho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Sakakura
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehirocho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Department of Physiology, School of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Numata
authors.affiliation: Department of Physiology, School of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Inoue
authors.affiliation: Department of Physiology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.|Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo 102-0075, Japan.
authors.forename: Michisuke
authors.identifier: 0000-0002-5750-3544
authors.initials: M
authors.lastname: Yuzaki
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.|Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo 102-0075, Japan.
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D000495:Allosteric Site; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002540:Cerebral Cortex / Q000378:metabolism; D056831:Coordination Complexes / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D018690:Excitatory Amino Acid Agonists / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D057809:HEK293 Cells; D006639:Histidine / Q000737:chemistry; D006801:Humans; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D010165:Palladium / Q000737:chemistry; D010766:Phosphorylation; D017207:Rats, Sprague-Dawley; D017470:Receptors, Glutamate / Q000235:genetics / Q000378:metabolism*; D058468:Receptors, Ionotropic Glutamate / Q000235:genetics / Q000378:metabolism*; D018094:Receptors, Metabotropic Glutamate / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C115163:CREB1 protein, human; D056831:Coordination Complexes; D017362:Cyclic AMP Response Element-Binding Protein; D018690:Excitatory Amino Acid Agonists; D017470:Receptors, Glutamate; D058468:Receptors, Ionotropic Glutamate; D018094:Receptors, Metabotropic Glutamate; C492973:alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid subtype glutamate receptor, human; C104077:metabotropic glutamate receptor type 1; D006639:Histidine; D010165:Palladium; D002118:Calcium