Versatile and Efficient Site-Specific Protein Functionalization by Tubulin Tyrosine Ligase.
A novel chemoenzymatic approach for simple and fast site-specific protein labeling is reported. Recombinant tubulin tyrosine ligase (TTL) was repurposed to attach various unnatural tyrosine derivatives as small bioorthogonal handles to proteins containing a short tubulin-derived recognition sequence (Tub-tag). This novel strategy enables a broad range of high-yielding and fast chemoselective C-terminal protein modifications on isolated proteins or in cell lysates for applications in biochemistry, cell biology, and beyond, as demonstrated by the site-specific labeling of nanobodies, GFP, and ubiquitin.
['Schumacher|Dominik|D|', 'Helma|Jonas|J|', 'Mann|Florian A|FA|', 'Pichler|Garwin|G|', 'Natale|Francesco|F|', 'Krause|Eberhard|E|', 'Cardoso|M Cristina|MC|', 'Hackenberger|Christian P R|CP|', 'Leonhardt|Heinrich|H|']
D006367:HeLa Cells D006801:Humans D008958:Models, Molecular D015394:Molecular Structure D010453:Peptide Synthases D011994:Recombinant Proteins D014443:Tyrosine
tubulin tyrosine ligase nanobodies antibodies chemoenzymatic labeling site-specific functionalization