title: Site-Specific Glycoconjugation of Protein via Bioorthogonal Tetrazine Cycloaddition with a Genetically Encoded trans-Cyclooctene or Bicyclononyne.
abstract: Efficient access to proteins modified site-specifically with glycans is important in glycobiology and for therapeutic applications. Herein, we report a biocompatible protein glycoconjugation by inverse demand Diels-Alder reaction between tetrazine and trans-cyclooctene. Tetrazine functionalized glycans were obtained in one step by CuAAC (Cu-catalyzed alkyne azide cycloaddition) between glycosyl azide and an alkyne-tetrazine adduct. Site-specific glycoconjugation was performed chemoselectively on a target protein in which a trans-cyclooctene derivatized lysine was genetically encoded. Glycoconjugation proceeded to completion on purified protein and was shown to be selective for the target protein in E. coli.
authors.affiliation: †Department of Organic Chemistry, NCCR Chemical Biology, University of Geneva, 30 quai Ernest Ansermet, 1211 Geneva, Switzerland.
authors.forename: Takuya
authors.initials: T
authors.lastname: Machida
authors.affiliation: ‡Technical University Munich, Institute for Advanced Study, Department of Chemistry, 4 Lichtenbergstraße, 85748 Garching, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Lang
authors.affiliation: §Ecole Polytechnique Fédérale de Lausanne (EPFL), Institut des sciences et ingénierie chimiques, NCCR Chemical Biology, 1015 Lausanne, Switzerland.
authors.forename: Lin
authors.initials: L
authors.lastname: Xue
authors.affiliation: ⊥Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0GH, United Kingdom.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
authors.affiliation: †Department of Organic Chemistry, NCCR Chemical Biology, University of Geneva, 30 quai Ernest Ansermet, 1211 Geneva, Switzerland.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Winssinger
mesh_terms: D000480:Alkynes; D001386:Azides / Q000737:chemistry; D001665:Binding Sites; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D061565:Cycloaddition Reaction; D034242:Cyclooctanes / Q000737:chemistry*; D003516:Cycloparaffins / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics*; D008958:Models, Molecular; D011487:Protein Conformation; D013379:Substrate Specificity
chemical_list: D000480:Alkynes; D001386:Azides; D034242:Cyclooctanes; D003516:Cycloparaffins; D006023:Glycoproteins; D003300:Copper