title: Incorporation of Amino Acids with Long-Chain Terminal Olefins into Proteins.
abstract: 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.
authors.affiliation: Institute of Chemistry, Technische Universität Berlin, Mueller-Breslau-Strasse 10, 10623 Berlin, Germany. m.p.exner@gmx.net.
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Exner
authors.affiliation: Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany. koehling@zedat.fu-berlin.de.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Köhling
authors.affiliation: Service de Chimie Bioorganique et de Marquage, iBiTecS, CEA, 91191 Gif-sur-Yvette, France. julie.rivollier@cea.fr.
authors.forename: Julie
authors.initials: J
authors.lastname: Rivollier
authors.affiliation: Service de Chimie Bioorganique et de Marquage, iBiTecS, CEA, 91191 Gif-sur-Yvette, France. sgosling@isthmus.fr.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Gosling
authors.affiliation: Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium. puneet.srivastava@rega.kuleuven.be.
authors.forename: Puneet
authors.initials: P
authors.lastname: Srivastava
authors.affiliation: Institute of Chemistry, Technische Universität Berlin, Mueller-Breslau-Strasse 10, 10623 Berlin, Germany. zheni.ivanova@gmail.com.
authors.forename: Zheni I
authors.initials: ZI
authors.lastname: Palyancheva
authors.affiliation: Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium. piet.herdewijn@rega.kuleuven.be.
authors.forename: Piet
authors.initials: P
authors.lastname: Herdewijn
authors.affiliation: Service de Chimie Bioorganique et de Marquage, iBiTecS, CEA, 91191 Gif-sur-Yvette, France. marie-pierre.heck@cea.fr.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Heck
authors.affiliation: Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany. joerg.rademann@fu-berlin.de.
authors.forename: Jörg
authors.initials: J
authors.lastname: Rademann
authors.affiliation: Institute of Chemistry, Technische Universität Berlin, Mueller-Breslau-Strasse 10, 10623 Berlin, Germany. budisa@biocat.tu-berlin.de.
authors.forename: Nediljko
authors.initials: N
authors.lastname: Budisa
mesh_terms: D000475:Alkenes / Q000737:chemistry*; D000596:Amino Acids / Q000737:chemistry*; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D008958:Models, Molecular; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D000475:Alkenes; D000596:Amino Acids; D011506:Proteins; D000604:Amino Acyl-tRNA Synthetases
keywords: metathesis; noncanonical amino acid; protein decoration; pyrrolysyl-tRNA synthetase; stop codon suppression; thiol-ene coupling