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dc.contributor.author | Tajima, Shunsuke | en |
dc.contributor.author | Nakata, Eiji | en |
dc.contributor.author | Sakaguchi, Reiko | en |
dc.contributor.author | Saimura, Masayuki | en |
dc.contributor.author | Mori, Yasuo | en |
dc.contributor.author | Morii, Takashi | en |
dc.contributor.alternative | 田嶋, 竣介 | ja |
dc.contributor.alternative | 中田, 栄司 | ja |
dc.contributor.alternative | 才村, 正幸 | ja |
dc.contributor.alternative | 森井, 孝 | ja |
dc.date.accessioned | 2022-07-04T08:48:33Z | - |
dc.date.available | 2022-07-04T08:48:33Z | - |
dc.date.issued | 2022-05-20 | - |
dc.identifier.uri | http://hdl.handle.net/2433/274689 | - |
dc.description.abstract | Auto-fluorescent protein (AFP)-based biosensors transduce the structural change in their embedded recognition modules induced by recognition/reaction events to fluorescence signal changes of AFP. The lack of detailed structural information on the recognition module often makes it difficult to optimize AFP-based biosensors. To enhance the signal response derived from detecting the putative structural change in the nitric oxide (NO)-sensing segment of transient receptor potential canonical 5 (TRPC5) fused to enhanced green fluorescent protein (EGFP), EGFP-TRPC5, a facile two-step screening strategy, in silico first and in vitro second, was applied to variants of EGFP-TRPC5 deletion-mutated within the recognition module. In in silico screening, the structural changes of the recognition modules were evaluated as root-mean-square-deviation (RMSD) values, and 10 candidates were efficiently selected from 47 derivatives. Through in vitro screening, four mutants were identified that showed a larger change in signal response than the parent EGFP-TRPC5. One mutant in particular, 551-575, showed four times larger change upon reaction with NO and H₂O₂. Furthermore, mutant 551-575 also showed a signal response upon reaction with H₂O₂ in mammalian HEK293 cells, indicating that the mutant has the potential to be applied as a biosensor for cell measurement. Therefore, this two-step screening method effectively allows the selection of AFP-based biosensors with sufficiently enhanced signal responses for application in mammalian cells. | en |
dc.language.iso | eng | - |
dc.publisher | Royal Society of Chemistry (RSC) | en |
dc.rights | © The Royal Society of Chemistry 2022 | en |
dc.rights | This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. | en |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/3.0/ | - |
dc.title | A two-step screening to optimize the signal response of an auto-fluorescent protein-based biosensor | en |
dc.type | journal article | - |
dc.type.niitype | Journal Article | - |
dc.identifier.jtitle | RSC Advances | en |
dc.identifier.volume | 12 | - |
dc.identifier.issue | 24 | - |
dc.identifier.spage | 15407 | - |
dc.identifier.epage | 15419 | - |
dc.relation.doi | 10.1039/d2ra02226e | - |
dc.textversion | publisher | - |
dc.identifier.pmid | 35693243 | - |
dcterms.accessRights | open access | - |
datacite.awardNumber | 17H05529 | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PUBLICLY-17H05529/ | - |
dc.identifier.eissn | 2046-2069 | - |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.awardTitle | 活性酸素種による翻訳後修飾を検出する蛍光バイオセンサー | ja |
出現コレクション: | 学術雑誌掲載論文等 |
このアイテムは次のライセンスが設定されています: クリエイティブ・コモンズ・ライセンス