title: A Versatile Approach for Site-Specific Lysine Acylation in Proteins.
abstract: Using amber suppression in coordination with a mutant pyrrolysyl-tRNA synthetase-tRNAPylpair, azidonorleucine is genetically encoded in E. coli. Its genetic incorporation followed by traceless Staudinger ligation with a phosphinothioester allows the convenient synthesis of a protein with a site-specifically installed lysine acylation. By simply changing the phosphinothioester identity, any lysine acylation type could be introduced. Using this approach, we demonstrated that both lysine acetylation and lysine succinylation can be installed selectively in ubiquitin and synthesized histone H3 with succinylation at its K4 position (H3K4su). Using an H3K4su-H4 tetramer as a substrate, we further confirmed that Sirt5 is an active histone desuccinylase. Lysine succinylation is a recently identified post-translational modification. The reported technique makes it possible to explicate regulatory functions of this modification in proteins.
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Zhipeng A
authors.initials: ZA
authors.lastname: Wang
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Yadagiri
authors.initials: Y
authors.lastname: Kurra
authors.affiliation: Department of Plant Pathology and Microbiology, Institute for Plant Genomics, Office of the Texas State Chemist, Department of Veterinary Pathobiology, College Station, TX, 77843, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Yan-Jiun
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Vangmayee
authors.initials: V
authors.lastname: Sharma
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY, 14853, USA.
authors.forename: Hening
authors.initials: H
authors.lastname: Lin
authors.affiliation: Department of Plant Pathology and Microbiology, Institute for Plant Genomics, Office of the Texas State Chemist, Department of Veterinary Pathobiology, College Station, TX, 77843, USA.
authors.forename: Susie Y
authors.initials: SY
authors.lastname: Dai
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Wenshe R
authors.identifier: 0000-0002-7078-6534
authors.initials: WR
authors.lastname: Liu
mesh_terms: D000215:Acylation; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics; D001386:Azides; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D006657:Histones / Q000235:genetics; D008239:Lysine / Q000032:analysis* / Q000235:genetics; D009646:Norleucine / Q000031:analogs & derivatives / Q000235:genetics; D011499:Protein Processing, Post-Translational
chemical_list: D001386:Azides; D029968:Escherichia coli Proteins; D006657:Histones; C000609218:azidonorleucine; D009646:Norleucine; D000604:Amino Acyl-tRNA Synthetases; D008239:Lysine
keywords: amber suppression; azidonorleucine; lysine acylation; protein modification; traceless Staudinger ligation