title: In-cell covalent labeling of reactive His-tag fused proteins.
abstract: A new method for in-cell protein labeling was developed. This method employed a binding-induced nucleophilic reaction between the Cys-appended His-tag and the Ni(II)-NTA containing an α-chloroacetamide. Using this method, not only labeling of His-tag fused proteins but also the detection of a protein-protein interaction was achieved inside living cells.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Kyoto University, Katsura, Kyoto 615-8510, Japan.
authors.forename: Shohei
authors.initials: S
authors.lastname: Uchinomiya
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nonaka
authors.forename: Sho
authors.initials: S
authors.lastname: Wakayama
authors.forename: Akio
authors.initials: A
authors.lastname: Ojida
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D000081:Acetamides / Q000737:chemistry; D045744:Cell Line, Tumor; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006639:Histidine / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009532:Nickel / Q000737:chemistry; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D060066:Protein Interaction Maps; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D022061:Tacrolimus Binding Protein 1A / Q000235:genetics / Q000378:metabolism
chemical_list: D000081:Acetamides; D005456:Fluorescent Dyes; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C013874:chloroacetamide; D006639:Histidine; D009532:Nickel; D022061:Tacrolimus Binding Protein 1A; D003545:Cysteine