title: Genetic code expansion enabled site-specific dual-color protein labeling: superresolution microscopy and beyond.
abstract: Genetic code expansion is emerging as an important tool for manipulation and labeling of proteins in vitro and in vivo. In combination with click-chemistry it allows site-specific labeling of target proteins with small organic fluorophores. This is achieved by cotranslational incorporation of noncanonical amino acids (ncAAs) in target proteins via orthogonal tRNA/amino-acyl tRNA synthetase pairs. In a subsequent step, ncAAs are labeled with small dyes via click-chemistry. Small labeling tags and free choice of which fluorophore to use and where to put it into the protein are of particular importance for single molecule science and superresolution microscopy. Such genetically encoded click chemistry tools facilitate high resolution studies of protein function in live cells, viral imaging, as well as the improved design of antibody-drug conjugates.
authors: Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Ivana
authors: I
authors: Nikić
authors: Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: lemke@embl.de.
authors: Edward A
authors: EA
authors: Lemke
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D057930:Click Chemistry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D005815:Genetic Code; D006801:Humans; D018796:Immunoconjugates / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D008958:Models, Molecular; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000596:Amino Acids; D005456:Fluorescent Dyes; D018796:Immunoconjugates; D011506:Proteins; D014764:Viral Proteins