title: Proteomic analysis of fatty-acylated proteins in mammalian cells with chemical reporters reveals S-acylation of histone H3 variants.
abstract: Bioorthogonal chemical reporters are useful tools for visualizing and identifying post-translational modifications on proteins. Here we report the proteomic analysis of mammalian proteins targeted by a series of fatty acid chemical reporters ranging from myristic to stearic acid. The large-scale analysis of total cell lysates from fully solubilized Jurkat T cells identified known fatty-acylated proteins and many new candidates, including nuclear proteins and in particular histone H3 variants. We demonstrate that histones H3.1, H3.2, and H3.3 are modified with fatty acid chemical reporters and identify the conserved cysteine 110 as a new site of S-acylation on histone H3.2. This newly discovered modification of histone H3 could have implications for nuclear organization and chromatin regulation. The unbiased proteomic analysis of fatty-acylated proteins using chemical reporters has revealed a greater diversity of lipid-modified proteins in mammalian cells and identified a novel post-translational modification of histones.
authors.affiliation: The Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, NY 10065, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Wilson
authors.forename: Anuradha S
authors.initials: AS
authors.lastname: Raghavan
authors.forename: Yu-Ying
authors.initials: YY
authors.lastname: Yang
authors.forename: Guillaume
authors.initials: G
authors.lastname: Charron
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
mesh_terms: D000215:Acylation; D000818:Animals; D057930:Click Chemistry; D003300:Copper / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D019169:Jurkat Cells; D008322:Mammals / Q000378:metabolism*; D015335:Molecular Probes / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D005227:Fatty Acids; D006657:Histones; D015335:Molecular Probes; D020033:Protein Isoforms; D020543:Proteome; D003300:Copper