title: Use of synthetic peptides for identifying biotinylation sites in human histones.
abstract: Posttranslational modifications of histones play an important role in the regulation of chromatin structure and, hence, gene regulation. Recently, we have identified a novel modification of histones: binding of the vitamin biotin to lysine residues in histones H2A, H3, and H4. Here, we describe a procedure to identify those amino acids that are targets for biotinylation in histones. Briefly, the following analytical sequence is used to identify biotinylation sites: (i) short peptides (<20 amino acids in length) are synthesized chemically; amino acid sequences in the peptides are based on the sequence in a given region of a given histone; (ii) peptides are incubated with biotinidase or holocarboxylase synthetase to conduct enzymatic biotinylation; and (iii) biotin in peptides are probed using streptavidin peroxidase. Amino acid substitutions (e.g., lysine-to-alanine substitutions) in synthetic peptides can be used to corroborate identification of biotinylation sites.
authors.affiliation: Department of Nutrition and Health Sciences, University of Nebraska at Lincoln, Lincoln, NE, USA.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Camporeale
authors.forename: Yap Ching
authors.initials: YC
authors.lastname: Chew
authors.forename: Alice
authors.initials: A
authors.lastname: Kueh
authors.forename: Gautam
authors.initials: G
authors.lastname: Sarath
authors.forename: Janos
authors.initials: J
authors.lastname: Zempleni
mesh_terms: D000818:Animals; D043523:Biotinidase / Q000097:blood / Q000378:metabolism; D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism; D006657:Histones / Q000737:chemistry*; D006801:Humans; D008099:Liver / Q000201:enzymology; D051379:Mice; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D013154:Spleen / Q000201:enzymology; D019809:Streptavidin / Q000737:chemistry
chemical_list: D006657:Histones; D010455:Peptides; D019809:Streptavidin; D043523:Biotinidase; D019731:Carbon-Nitrogen Ligases; C023312:holocarboxylase synthetases