Ethynylation of Cysteines from Peptides to Proteins in Living Cells.
Fiche publication
Date publication
mars 2020
Journal
Angewandte Chemie (International ed. in English)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CIANFERANI Sarah, Dr WAGNER Alain, Dr CHAUBET Guilhem
Tous les auteurs :
Tessier R, Nandi RK, Dwyer BG, Abegg D, Sornay C, Ceballos J, Erb S, Cianferani S, Wagner A, Chaubet G, Adibekian A, Waser J
Lien Pubmed
Résumé
Efficient methods to introduce bioorthogonal groups, such as terminal alkynes, into biomolecules are important tools for chemical biology. State-of-the-art approaches are based on the introduction of a linker between the targeted amino acid and the alkyne, and still present limitations of either reactivity, selectivity or adduct stability. Herein, we present an ethynylation method of cysteine residues based on the use of ethynylbenziodoxolone (EBX) reagents. In contrast to other approaches, the acetylene group is directly introduced onto the thiol group of cysteine and can be used in one-pot in a copper-catalyzed alkyne-azide cycloaddition (CuAAC) for further functionalization. Labeling proceeded with reaction rates comparable or higher than the most often used iodoacetamide on peptides or maleimide on the antibody trastuzumab. Under optimized conditions, high cysteine selectivity was observed. The reagents were also used in living cells for cysteine proteomic profiling and displayed an improved coverage of the cysteinome compared to previously reported iodoacetamide or hypervalent iodine-reagent based probes. Fine-tuning of the EBX reagents allowed optimization of their reactivity and physical properties for the desired application.
Mots clés
Alkynes, Hypervalent Iodine Reagents, bioconjugation, cysteine proteomic, peptide and antibody functionalization
Référence
Angew. Chem. Int. Ed. Engl.. 2020 Mar 31;: