Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells

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...

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Bibliographic Details
Published in:Nature chemistry 2016-10, Vol.8 (10), p.958-967
Main Authors: Kiyonaka, Shigeki, Kubota, Ryou, Michibata, Yukiko, Sakakura, Masayoshi, Takahashi, Hideo, Numata, Tomohiro, Inoue, Ryuji, Yuzaki, Michisuke, Hamachi, Itaru
Format: Article
Language:English
Subjects:
Allosteric Site
Animals
Binding sites
Calcium - metabolism
Cerebral Cortex - metabolism
Chemistry
Coordination Complexes - chemistry
Coordination Complexes - pharmacology
Coordination Complexes - toxicity
Cyclic AMP Response Element-Binding Protein - metabolism
Enzymes
Excitatory Amino Acid Agonists - chemistry
Excitatory Amino Acid Agonists - pharmacology
Excitatory Amino Acid Agonists - toxicity
HEK293 Cells
Histidine - chemistry
Humans
Ligands
Mutation
Neurons - metabolism
Palladium - chemistry
Phosphorylation
Physiology
Proteins
Rats, Sprague-Dawley
Receptors, Glutamate - genetics
Receptors, Glutamate - metabolism
Receptors, Ionotropic Glutamate - genetics
Receptors, Ionotropic Glutamate - metabolism
Receptors, Metabotropic Glutamate - genetics
Receptors, Metabotropic Glutamate - metabolism
Signal Transduction
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Summary: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.
ISSN:1755-4330
1755-4349
DOI:10.1038/nchem.2554