Design, Synthesis and Catalytic Activity of Protein Containing Thiotyrosine as an Active Site Residue.
Fiche publication
Date publication
avril 2024
Journal
Chembiochem : a European journal of chemical biology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr KIEFFER Bruno
Tous les auteurs :
Bachelart T, Kumar S, Jouin A, Yousef M, Kieffer B, Torbeev V
Lien Pubmed
Résumé
Native chemical ligation is a key reaction in the toolbox of chemical methods for the synthesis of native and modified proteins. The catalysis of ligation is commonly performed by using small aryl-thiol molecules added at high concentrations. In this work, we incorporated thiotyrosine, a non-canonical amino acid containing an aryl-thiol moiety, into a designed cyclic protein « sans queue ni tête ». Importantly, the protein environment reduced the pKa of the thiol group to 5.8-5.9, which is significantly lower than the previously reported value for thiotyrosine in a short peptide (pKa6.4). Furthermore, we demonstrated the catalytic activity of this protein both as hydrolase and in native chemical ligation of peptides. These results will be useful for the development of efficient protein catalysts (enzymes) for protein synthesis and modification.
Mots clés
aryl-thiols, biocatalysis, chemical protein synthesis, cyclic proteins, protein design
Référence
Chembiochem. 2024 04 17;:e202400148