title: Versatile and Efficient Site-Specific Protein Functionalization by Tubulin Tyrosine Ligase.
abstract: 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.
authors.affiliation: Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Berlin (Germany).|Department of Chemistry, Humboldt Universität zu Berlin, Berlin (Germany).
authors.forename: Dominik
authors.initials: D
authors.lastname: Schumacher
authors.affiliation: Department of Biology II, Ludwig Maximilians Universität München and Center for Integrated Protein Science Munich, Martinsried (Germany).
authors.forename: Jonas
authors.initials: J
authors.lastname: Helma
authors.affiliation: Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Berlin (Germany).|Department of Chemistry, Humboldt Universität zu Berlin, Berlin (Germany).
authors.forename: Florian A
authors.initials: FA
authors.lastname: Mann
authors.affiliation: Department of Biology II, Ludwig Maximilians Universität München and Center for Integrated Protein Science Munich, Martinsried (Germany).
authors.forename: Garwin
authors.initials: G
authors.lastname: Pichler
authors.affiliation: Department of Biology, Technische Universität Darmstadt, Darmstadt (Germany).
authors.forename: Francesco
authors.initials: F
authors.lastname: Natale
authors.affiliation: Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Berlin (Germany).
authors.forename: Eberhard
authors.initials: E
authors.lastname: Krause
authors.affiliation: Department of Biology, Technische Universität Darmstadt, Darmstadt (Germany).
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Cardoso
authors.affiliation: Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Berlin (Germany). hackenbe@fmp-berlin.de.|Department of Chemistry, Humboldt Universität zu Berlin, Berlin (Germany). hackenbe@fmp-berlin.de.
authors.forename: Christian P R
authors.initials: CP
authors.lastname: Hackenberger
authors.affiliation: Department of Biology II, Ludwig Maximilians Universität München and Center for Integrated Protein Science Munich, Martinsried (Germany). h.leonhardt@lmu.de.
authors.forename: Heinrich
authors.initials: H
authors.lastname: Leonhardt
mesh_terms: D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D010453:Peptide Synthases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014443:Tyrosine; D010453:Peptide Synthases; C023213:tyrosyltubulin ligase
keywords: antibodies; chemoenzymatic labeling; nanobodies; site-specific functionalization; tubulin tyrosine ligase