Site-Specific Turn-On Fluorescent Labeling of DNA-Interacting Protein Using Oligodeoxynucleotides That Modify Lysines to Produce 5,6-Dimethoxy 3-Methyleneisoindolin-1-one

Chiemi Gatanaga, Bo Yang, Yuka Inadomi, Kazuteru Usui, Chiyoe Ota, Tsutomu Katayama, Hiroshi Suemune, Mariko Aso

研究成果: ジャーナルへの寄稿記事

4 引用 (Scopus)

抄録

We have developed oligodeoxynucleotides (ODNs) that modify primary amines to produce 5,6-dimethoxy 3-methyleneisoindolin-1-one. Compared to the oxygen isosteric fluorophore, 4,5-dimethoxyphthalimide, this methyleneisoindolinone was more stable and exhibited an 85 nm blue-shifted fluorescent emission (λmax at 425 nm) with an intensity comparable to that of the phthalimide. Reaction of the DNA-binding domain of Escherichia coli DnaA protein with an ODN containing its binding sequence efficiently afforded a modified fluorescent protein at a specific lysine residue in the proximity of the ODN. A full-length DnaA protein was also successfully fluorescently labeled. These results demonstrate the potential utility of the ODNs developed in this study for the fluorescent labeling of DNA-interacting protein at the lysine residue of interest.

元の言語英語
ページ(範囲)2216-2221
ページ数6
ジャーナルACS Chemical Biology
11
発行部数8
DOI
出版物ステータス出版済み - 8 19 2016

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Oligodeoxyribonucleotides
Labeling
Lysine
DNA
Proteins
Fluorophores
Escherichia coli Proteins
Escherichia coli
Amines
Oxygen
3-methyleneisoindolin-1-one

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine

これを引用

Site-Specific Turn-On Fluorescent Labeling of DNA-Interacting Protein Using Oligodeoxynucleotides That Modify Lysines to Produce 5,6-Dimethoxy 3-Methyleneisoindolin-1-one. / Gatanaga, Chiemi; Yang, Bo; Inadomi, Yuka; Usui, Kazuteru; Ota, Chiyoe; Katayama, Tsutomu; Suemune, Hiroshi; Aso, Mariko.

:: ACS Chemical Biology, 巻 11, 番号 8, 19.08.2016, p. 2216-2221.

研究成果: ジャーナルへの寄稿記事

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abstract = "We have developed oligodeoxynucleotides (ODNs) that modify primary amines to produce 5,6-dimethoxy 3-methyleneisoindolin-1-one. Compared to the oxygen isosteric fluorophore, 4,5-dimethoxyphthalimide, this methyleneisoindolinone was more stable and exhibited an 85 nm blue-shifted fluorescent emission (λmax at 425 nm) with an intensity comparable to that of the phthalimide. Reaction of the DNA-binding domain of Escherichia coli DnaA protein with an ODN containing its binding sequence efficiently afforded a modified fluorescent protein at a specific lysine residue in the proximity of the ODN. A full-length DnaA protein was also successfully fluorescently labeled. These results demonstrate the potential utility of the ODNs developed in this study for the fluorescent labeling of DNA-interacting protein at the lysine residue of interest.",
author = "Chiemi Gatanaga and Bo Yang and Yuka Inadomi and Kazuteru Usui and Chiyoe Ota and Tsutomu Katayama and Hiroshi Suemune and Mariko Aso",
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AU - Gatanaga, Chiemi

AU - Yang, Bo

AU - Inadomi, Yuka

AU - Usui, Kazuteru

AU - Ota, Chiyoe

AU - Katayama, Tsutomu

AU - Suemune, Hiroshi

AU - Aso, Mariko

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AB - We have developed oligodeoxynucleotides (ODNs) that modify primary amines to produce 5,6-dimethoxy 3-methyleneisoindolin-1-one. Compared to the oxygen isosteric fluorophore, 4,5-dimethoxyphthalimide, this methyleneisoindolinone was more stable and exhibited an 85 nm blue-shifted fluorescent emission (λmax at 425 nm) with an intensity comparable to that of the phthalimide. Reaction of the DNA-binding domain of Escherichia coli DnaA protein with an ODN containing its binding sequence efficiently afforded a modified fluorescent protein at a specific lysine residue in the proximity of the ODN. A full-length DnaA protein was also successfully fluorescently labeled. These results demonstrate the potential utility of the ODNs developed in this study for the fluorescent labeling of DNA-interacting protein at the lysine residue of interest.

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