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Photocycle features of heterologously expressed and assembled eukaryotic flavin-binding BLUF domains of photoactivated adenylyl cyclase (PAC), a blue-light receptor in Euglena gracilis
https://ir.soken.ac.jp/records/3884
https://ir.soken.ac.jp/records/388412214d1a-03e5-4f89-a04b-6c27931da792
名前 / ファイル | ライセンス | アクション |
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2013-08-13 | |||||
タイトル | ||||||
タイトル | Photocycle features of heterologously expressed and assembled eukaryotic flavin-binding BLUF domains of photoactivated adenylyl cyclase (PAC), a blue-light receptor in Euglena gracilis | |||||
タイトル | ||||||
タイトル | Photocycle features of heterologously expressed and assembled eukaryotic flavin-binding BLUF domains of photoactivated adenylyl cyclase (PAC), a blue-light receptor in Euglena gracilis | |||||
言語 | en | |||||
言語 | ||||||
言語 | eng | |||||
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資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
ITO, Shinji
× ITO, Shinji× MURAKAMI, Akio× SATO, Kyosuke× NISHINA, Yasuzo× SHIGA, Kiyoshi× TAKAHASHI, Tetsuo× HIGASHI, Shoichi× ISEKI, Mineo× WATANABE, Masakatsu |
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著者別名 |
渡辺, 正勝
× 渡辺, 正勝 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Photoactivated adenylyl cyclase (PAC) is a recently discovered blue-light photoreceptor that mediates photomovement in Euglena gracilis (Iseki et al., Nature, 2002, 415, 1047-1051). PAC appears to be a heterotetramer composed of two FAD-binding subunits (PACα and PACβ). Both subunits have a pair of homologous regions (F1 and F2) which show homology with prokaryotic "sensors of blue-light using FAD" (BLUF) domains. The F1 and F2 domains of PAC are the only eukaryotic BLUF domains found thus far. We obtained soluble recombinant F1 and F2 proteins in PACα by heterologous expression with fused glutathione-S-transferase (GST) in E. coli. The expressed F1 samples did not bind flavins, but the F2 samples contained both FAD and FMN with trace amounts of riboflavin. We also assembled the histidine-tagged recombinant F2 (6His-F2) from inclusion bodies in E. coli with exogenous FAD or FMN. Blue-light-induced changes in absorption spectra of these assembled samples were highly similar to those reported for prokaryotic BLUF domains. The FAD- or FMN-assembled 6His-F2 photocycled with nearly the same rate constants of light-reaction and dark-relaxation, which were slightly lower than those of GST-cleaved F2. The estimated quantum efficiency for the phototransformation was 0.28-0.32, and the half-life was 34-44 s at 25 °C for the recombinant PACα F2, whereas that reported for prokaryotic BLUF domains varied from ca. 3.5 s (Tl10078) to ca. 900 s (AppA). The mutated recombinant Y472F and Q514G of PACα F2 and the F2 domain of the PACα homologue from Eutreptiella gymnastica, which lacks the Gln residue conserved in other BLUF domains, showed no photoinduced transformation. | |||||
書誌情報 |
Photochemical and Photobiological Sciences en : Photochemical and Photobiological Sciences 巻 4, 号 9, p. 762-769, 発行日 2005-09 |
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出版者 | ||||||
出版者 | Royal Society of Chemistry | |||||
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収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1474905X | |||||
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関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | 16121289 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | http://doi.org/10.1039/b505792b | |||||
関連名称 | 10.1039/b505792b | |||||
権利 | ||||||
権利情報 | © The Royal Society of Chemistry and Owner Societies 2005 | |||||
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識別子タイプ | URI | |||||
関連識別子 | http://www.rsc.org/AboutUs/Copyright/LicencetoPublishforjournals.asp | |||||
関連名称 | Copyright | |||||
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内容記述タイプ | Other | |||||
内容記述 | application/pdf | |||||
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出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 |