title: Site-Specific Turn-On Fluorescent Labeling of DNA-Interacting Protein Using Oligodeoxynucleotides That Modify Lysines To Produce 5,6-Dimethoxy 3-Methyleneisoindolin-1-one.
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.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Chiemi
authors.initials: C
authors.lastname: Gatanaga
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Bo
authors.initials: B
authors.lastname: Yang
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Inadomi
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Kazuteru
authors.initials: K
authors.lastname: Usui
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Chiyoe
authors.initials: C
authors.lastname: Ota
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Katayama
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suemune
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Mariko
authors.initials: M
authors.lastname: Aso
mesh_terms: D004268:DNA-Binding Proteins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D054833:Isoindoles / Q000138:chemical synthesis*; D008239:Lysine / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
chemical_list: D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D054833:Isoindoles; D009838:Oligodeoxyribonucleotides; D008239:Lysine