title: Traceless semisynthesis of a set of histone 3 species bearing specific lysine methylation marks.
abstract: Considerable mechanistic insight into the function of histone post-translational modifications and the enzymes that install and remove them derives from in vitro experiments with modified histones, often embedded in nucleosomes. We report the first semisyntheses of native-like histone 3 (H3) bearing tri- and dimethyllysines at position 79 and trimethyllysine at position 36, as well as more facile and traceless semisyntheses of K9 and K27 trimethylated species. These semisyntheses are practical on a multi-milligram scale and can also generate H3 with combinations of marks. Each of these modifications has distinct functional consequences, although the pathways by which H3K36me3 and H3K79me2/3 act have not been entirely mapped. To this end, we demonstrated that our semisynthetic histones, when reconstituted into nucleosomes, are valuable affinity reagents for unbiased binding partner discovery and compare them to their methyllysine analogue (MLA) counterparts at the nucleosome level.
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, 920 E. 58th St, Chicago, IL 60637 (USA).
authors.forename: Zhonglei
authors.initials: Z
authors.lastname: Chen
authors.forename: Adrian T
authors.initials: AT
authors.lastname: Grzybowski
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Ruthenburg
mesh_terms: D000595:Amino Acid Sequence; D006367:HeLa Cells; D006657:Histones / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D008745:Methylation; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational
chemical_list: D006657:Histones; D009707:Nucleosomes; C003712:trimethyllysine; D008239:Lysine
keywords: binding partner discovery; bioorganic chemistry; histone methylation; native chemical ligation; protein engineering