title: Ligand-directed tosyl chemistry for selective native protein labeling in vitro, in cells, and in vivo.
abstract: Introducing nongenetically encoded, synthetic probes into specific proteins is now recognized as a key component in chemical biology. In particular, the ability to chemically modify specific "native" proteins in various contexts from in vitro to cellular systems is of fundamental importance to study biological systems. We developed a protein-labeling technique termed ligand-directed tosyl (LDT) chemistry for this purpose. This method is capable of labeling specific native proteins with diverse synthetic probes with high site specificity and target selectivity without compromising protein function. Here we describe the principle of the LDT chemistry and the protocol for selective chemical labeling of native carbonic anhydrase in vitro, in blood cells (ex vivo), and in living mice (in vivo).
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Tsukiji
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D000818:Animals; D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D008024:Ligands; D008297:Male; D008813:Mice, Inbred ICR; D013194:Staining and Labeling; D014105:Tosyl Compounds / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008024:Ligands; D014105:Tosyl Compounds; D024402:Carbonic Anhydrase II