title: Reactive group-embedded affinity labeling reagent for efficient intracellular protein labeling.
abstract: Affinity labeling of a target protein is a powerful method for chemical biology studies. However, it is still difficult to label intracellular proteins efficiently in living cells. We propose the novel design strategy of a reactive group-embedded affinity labeling reagent for efficient protein labeling. With FKBP12 as the model target protein, the ligand binding pocket-oriented labeling reagent could label intracellular protein, whereas protein surface-oriented reagent was ineffective for labeling in living cells, partially because of the intracellular protein fluctuation under the macromolecular crowding effects. These results provide new insight for efficient intracellular protein labeling.
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan; Precursory Research for Embryonic Science and Technology PREST, Science and Technology Agency, JST, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Takaoka
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Nukadzuka
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan. Electronic address: ueda@m.tohoku.ac.jp.
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000345:Affinity Labels / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D022061:Tacrolimus Binding Protein 1A / Q000032:analysis*
chemical_list: D000345:Affinity Labels; D022061:Tacrolimus Binding Protein 1A