title: A chemical biology route to site-specific authentic protein modifications.
abstract: Many essential biological processes are controlled by posttranslational protein modifications. The inability to synthetically attain the diversity enabled by these modifications limits functional studies of many proteins. We designed a three-step approach for installing authentic posttranslational modifications in recombinant proteins. We first use the established O-phosphoserine (Sep) orthogonal translation system to create a Sep-containing recombinant protein. The Sep residue is then dephosphorylated to dehydroalanine (Dha). Last, conjugate addition of alkyl iodides to Dha, promoted by zinc and copper, enables chemoselective carbon-carbon bond formation. To validate our approach, we produced histone H3, ubiquitin, and green fluorescent protein variants with site-specific modifications, including different methylations of H3K79. The methylated histones stimulate transcription through histone acetylation. This approach offers a powerful tool to engineer diverse designer proteins.
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Aerin
authors.initials: A
authors.lastname: Yang
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Sura
authors.initials: S
authors.lastname: Ha
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Jihye
authors.initials: J
authors.lastname: Ahn
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Rira
authors.initials: R
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Sungyoon
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Younghoon
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Jaehoon
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, USA.
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Hee-Yoon
authors.initials: HY
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
authors.forename: Hee-Sung
authors.initials: HS
authors.lastname: Park
mesh_terms: D000107:Acetylation; D000409:Alanine / Q000031:analogs & derivatives / Q000737:chemistry; D000818:Animals; D003300:Copper / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry; D006657:Histones / Q000096:biosynthesis / Q000737:chemistry; D007454:Iodides / Q000737:chemistry; D008745:Methylation; D010768:Phosphoserine / Q000737:chemistry; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry*; D025801:Ubiquitin / Q000096:biosynthesis / Q000737:chemistry; D029867:Xenopus Proteins / Q000096:biosynthesis / Q000737:chemistry; D014982:Xenopus laevis; D015032:Zinc / Q000737:chemistry
chemical_list: D006657:Histones; D007454:Iodides; D011994:Recombinant Proteins; D025801:Ubiquitin; D029867:Xenopus Proteins; D049452:Green Fluorescent Proteins; D010768:Phosphoserine; D003300:Copper; C015102:dehydroalanine; D015032:Zinc; D000409:Alanine