title: Genetically encoded 1,2-aminothiols facilitate rapid and site-specific protein labeling via a bio-orthogonal cyanobenzothiazole condensation.
abstract: We report evolved orthogonal pyrrolysyl-tRNA synthetase/tRNA(CUA) pairs that direct the efficient, site-specific incorporation of N(ε)-L-thiaprolyl-L-lysine, N(ε)-D-cysteinyl-L-lysine, and N(ε)-L-cysteinyl-L-lysine into recombinant proteins in Escherichia coli . We demonstrate that the unique 1,2-aminothiol introduced by our approach can be efficiently, rapidly, and specifically labeled via a cyanobenzothiazole condensation to quantitatively introduce biophysical probes into proteins. Moreover, we show that, in combination with cysteine labeling, this approach allows the dual labeling of proteins with distinct probes at two distinct, genetically defined sites.
authors: Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors: Duy P
authors: DP
authors: Nguyen
authors: Thomas
authors: T
authors: Elliott
authors: Matthew
authors: M
authors: Holt
authors: Tom W
authors: TW
authors: Muir
authors: Jason W
authors: JW
authors: Chin
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry* / Q000378:metabolism; D052160:Benzothiazoles / Q000737:chemistry*; D001665:Binding Sites; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry; D017020:Methanosarcina / Q000201:enzymology; D008958:Models, Molecular; D008968:Molecular Conformation; D013194:Staining and Labeling; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D052160:Benzothiazoles; D013438:Sulfhydryl Compounds; D000604:Amino Acyl-tRNA Synthetases; C456839:pyrrolysine; D008239:Lysine