Use of synthetic peptides for identifying biotinylation sites in human histones

Gabriela Camporeale, Ching Chew Yap, Alice Kueh, Gautam Sarath, Janos Zempleni

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationAvidin-Biotin Interactions - Methods and Applications
PublisherHumana Press
Pages139-148
Number of pages10
ISBN (Print)9781597455794
DOIs
StatePublished - Dec 15 2007

Publication series

NameMethods in Molecular Biology
Volume418
ISSN (Print)1064-3745

Keywords

  • Biotinidase
  • Biotinylation
  • Chromatin
  • Histones
  • Holocarboxylase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Camporeale, G., Yap, C. C., Kueh, A., Sarath, G., & Zempleni, J. (2007). Use of synthetic peptides for identifying biotinylation sites in human histones. In Avidin-Biotin Interactions - Methods and Applications (pp. 139-148). (Methods in Molecular Biology; Vol. 418). Humana Press. https://doi.org/10.1385/1-59745-579-2:139