{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1360004233149965056.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1021/acschembio.7b01042"}},{"identifier":{"@type":"URI","@value":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.7b01042"}},{"identifier":{"@type":"PMID","@value":"29437380"}}],"resourceType":"学術雑誌論文(journal article)","dc:title":[{"@value":"Ligand-Directed Chemistry of AMPA Receptors Confers Live-Cell Fluorescent Biosensors"}],"description":[{"notation":[{"@value":"AMPA-type glutamate receptors (AMPARs) mediate fast excitatory synaptic transmission in the central nervous system. Dysregulation of AMPAR function is associated with many kinds of neurological, neurodegenerative, and psychiatric disorders. As a result, molecules capable of controlling AMPAR functions are potential therapeutic agents. Fluorescent semisynthetic biosensors have attracted considerable interest for the discovery of ligands selectively acting on target proteins. Given the large protein complex formation of AMPARs in live cells, biosensors using full-length AMPARs retaining original functionality are ideal for drug screening. Here, we demonstrate that fluorophore-labeled AMPARs prepared by ligand-directed acyl imidazole chemistry can act as turn-on fluorescent biosensors for AMPAR ligands in living cells. These biosensors selectively detect orthosteric ligands of AMPARs among the glutamate receptor family. Notably, the dissociation constants of agonists and antagonists for AMPARs were determined in live cells, which revealed that the ligand-binding properties of AMPARs to agonists are largely different in living cells, compared with noncellular conditions. We also show that these sensors can be applied to detecting allosteric modulators or subunit-selective ligands of AMPARs. Thus, our protein-based biosensors can be useful for discovering pharmaceutical agents to treat AMPAR-related neurological disorders."}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1380004233149965056","@type":"Researcher","foaf:name":[{"@value":"Shigeki Kiyonaka"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380004233149964929","@type":"Researcher","foaf:name":[{"@value":"Seiji Sakamoto"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380004233149965058","@type":"Researcher","foaf:name":[{"@value":"Sho Wakayama"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380004233149965057","@type":"Researcher","foaf:name":[{"@value":"Yuma Morikawa"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380004233149964928","@type":"Researcher","foaf:name":[{"@value":"Muneo Tsujikawa"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, 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Hamachi"}],"jpcoar:affiliationName":[{"@value":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan"},{"@value":"CREST(Core Research for Evolutional Science and Technology, JST), Chiyodaku, Tokyo, 102-0075, Japan"}]}],"publication":{"publicationIdentifier":[{"@type":"PISSN","@value":"15548929"},{"@type":"EISSN","@value":"15548937"}],"prism:publicationName":[{"@value":"ACS Chemical Biology"}],"dc:publisher":[{"@value":"American Chemical Society (ACS)"}],"prism:publicationDate":"2018-02-13","prism:volume":"13","prism:number":"7","prism:startingPage":"1880","prism:endingPage":"1889"},"reviewed":"false","url":[{"@id":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.7b01042"}],"createdAt":"2018-02-13","modifiedAt":"2025-07-01","foaf:topic":[{"@id":"https://cir.nii.ac.jp/all?q=Biosensing%20Techniques","dc:title":"Biosensing Techniques"},{"@id":"https://cir.nii.ac.jp/all?q=Ligands","dc:title":"Ligands"},{"@id":"https://cir.nii.ac.jp/all?q=Fluorescence","dc:title":"Fluorescence"},{"@id":"https://cir.nii.ac.jp/all?q=HEK293%20Cells","dc:title":"HEK293 Cells"},{"@id":"https://cir.nii.ac.jp/all?q=Drug%20Discovery","dc:title":"Drug Discovery"},{"@id":"https://cir.nii.ac.jp/all?q=Humans","dc:title":"Humans"},{"@id":"https://cir.nii.ac.jp/all?q=Receptors,%20AMPA","dc:title":"Receptors, AMPA"},{"@id":"https://cir.nii.ac.jp/all?q=Fluorescent%20Dyes","dc:title":"Fluorescent Dyes"}],"project":[{"@id":"https://cir.nii.ac.jp/crid/1040000781959581312","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"17H06348"},{"@type":"JGN","@value":"JP17H06348"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PLANNED-17H06348/"}],"notation":[{"@language":"ja","@value":"分子夾雑下での生命分子の直接修飾／機能解析を実現する有機化学"},{"@language":"en","@value":"Organic chemistry-based approaches for biomolecules modification under multimolecular crowding biosystems"}]}],"relatedProduct":[{"@id":"https://cir.nii.ac.jp/crid/1050282677899136384","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["references"],"jpcoar:relatedTitle":[{"@language":"en","@value":"Chemical labelling for visualizing native AMPA receptors in live neurons"},{"@value":"Chemical labeling for visualizing native AMPA receptors in live neurons"},{"@value":"Hamachi, I. Chemical labelling for visualizing native AMPA receptors in live neurons"}]},{"@id":"https://cir.nii.ac.jp/crid/1050282813187240960","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@language":"en","@value":"Live-cell imaging of membrane proteins by a coiled-coil labeling method—Principles and applications"}]},{"@id":"https://cir.nii.ac.jp/crid/1050860786106164352","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@language":"en","@value":"Recent applications of N-acyl imidazole chemistry in chemical biology"},{"@value":"Recent applications of <i>N</i>-acyl imidazole chemistry in chemical biology"}]},{"@id":"https://cir.nii.ac.jp/crid/1360002216625012864","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Recent Progress in Design of Protein-Based Fluorescent Biosensors and Their 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Acid (AMPA) Binding Sites Reflect Membrane‐Dependent States of a Single Receptor"}]},{"@id":"https://cir.nii.ac.jp/crid/1361699995731655040","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Design, Synthesis, and Pharmacology of a Highly Subtype-Selective GluR1/2 Agonist, (<i>RS</i>)-2-Amino-3-(4-chloro-3-hydroxy-5-isoxazolyl)propionic Acid (Cl-HIBO)"}]},{"@id":"https://cir.nii.ac.jp/crid/1361699996141530752","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"X-ray structure, symmetry and mechanism of an AMPA-subtype glutamate receptor"}]},{"@id":"https://cir.nii.ac.jp/crid/1361981468381359232","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Agonist discrimination between AMPA receptor subtypes"}]},{"@id":"https://cir.nii.ac.jp/crid/1362262944459008768","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Physiological significance of high- and low-affinity agonist binding to neuronal and recombinant AMPA receptors"}]},{"@id":"https://cir.nii.ac.jp/crid/1362262944466815616","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Pharmacology of ionotropic glutamate receptors: A structural perspective"}]},{"@id":"https://cir.nii.ac.jp/crid/1362544418923195648","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Mechanisms for Activation and Antagonism of an AMPA-Sensitive Glutamate Receptor"}]},{"@id":"https://cir.nii.ac.jp/crid/1362544419419510784","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"A Semisynthetic Fluorescent Sensor Protein for Glutamate"}]},{"@id":"https://cir.nii.ac.jp/crid/1362544419807074944","@type":"Article","relationType":["references"],"jpcoar:relatedTitle":[{"@value":"Glutamate Receptor Ion Channels: Structure, Regulation, and 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