Incorporation of Amino Acids with Long-Chain Terminal Olefins into Proteins
Matthias P. Exner, Sebastian Köhling, Julie Rivollier, Sandrine Gosling, Puneet Srivastava, Zheni I. Palyancheva, Piet Herdewijn, Marie-Pierre Heck, Jörg Rademann, and Nediljko Budisa – 2016
The increasing need for site-specific protein decorations that mimic natural posttranslational modifications requires access to a variety of noncanonical amino acids with moieties enabling bioorthogonal conjugation chemistry. Here we present the incorporation of long-chain olefinic amino acids into model proteins with rational variants of pyrrolysyl-tRNA synthetase (PylRS). Nε-heptenoyl lysine was incorporated for the first time using the known promiscuous variant PylRS(Y306A/Y384F), and Nε-pentenoyl lysine was incorporated in significant yields with the novel variant PylRS(C348A/Y384F). This is the only example of rational modification at position C348 to enlarge the enzyme’s binding pocket. Furthermore, we demonstrate the feasibility of our chosen amino acids in the thiol-ene conjugation reaction with a thiolated polysaccharide.