title: Synthesis of non-linear protein dimers through a genetically encoded Thiol-ene reaction.
abstract: Site-specific incorporation of bioorthogonal unnatural amino acids into proteins provides a useful tool for the installation of specific functionalities that will allow for the labeling of proteins with virtually any probe. We demonstrate the genetic encoding of a set of alkene lysines using the orthogonal PylRS/PylTCUA pair in Escherichia coli. The installed double bond functionality was then applied in a photoinitiated thiol-ene reaction of the protein with a fluorescent thiol-bearing probe, as well as a cysteine residue of a second protein, showing the applicability of this approach in the formation of heterogeneous non-linear fused proteins.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Jessica
authors.initials: J
authors.lastname: Torres-Kolbus
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Chungjung
authors.initials: C
authors.lastname: Chou
authors.affiliation: Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Jihe
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina, United States of America; Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D000475:Alkenes / Q000378:metabolism*; D000818:Animals; D005456:Fluorescent Dyes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism; D055503:Protein Multimerization; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D000475:Alkenes; D005456:Fluorescent Dyes; D009211:Myoglobin; D013438:Sulfhydryl Compounds; D049452:Green Fluorescent Proteins; D009113:Muramidase; D008239:Lysine