title: FRET-based analysis of protein-nucleic acid interactions by genetically incorporating a fluorescent amino acid.
abstract: Protein-nucleic acid interaction is an important process in many biological phenomena. In this study, a fluorescence resonance energy transfer (FRET)-based protein-DNA binding assay has been developed, in which a fluorescent amino acid is genetically incorporated into a DNA-binding protein. A coumarin-containing amino acid was incorporated into a DNA-binding protein, and the mutant protein specifically produced a FRET signal upon binding to its cognate DNA labeled with a fluorophore. The protein-DNA binding affinity was then measured under equilibrium conditions. This method is advantageous for studying protein-nucleic acid interactions, because it is performed under equilibrium conditions, technically easy, and applicable to any nucleic acid-binding protein.
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea.
authors.forename: Hyojin
authors.initials: H
authors.lastname: Park
authors.forename: Heejin
authors.initials: H
authors.lastname: Kang
authors.forename: Wooseok
authors.initials: W
authors.lastname: Ko
authors.forename: Wonkoo
authors.initials: W
authors.lastname: Lee
authors.forename: Kyubong
authors.initials: K
authors.lastname: Jo
authors.forename: Hyun Soo
authors.initials: HS
authors.lastname: Lee
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D007700:Kinetics; D011485:Protein Binding; D013194:Staining and Labeling
chemical_list: D000596:Amino Acids; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D004247:DNA