title: Purification of specific chromatin loci for proteomic analysis.
abstract: Purification of small, native chromatin regions for proteomic identification of specifically bound proteins and histone posttranslational modifications is a powerful approach for studying mechanisms of chromosome metabolism. Here we detail a Chromatin Affinity Purification with Mass Spectrometry (ChAP-MS) approach for affinity purification of 1 kb regions of chromatin for targeted proteomic analysis. This approach utilizes quantitative, high resolution mass spectrometry to categorize proteins and histone posttranslational modifications co-enriched with the given chromatin region as either "specific" to the targeted chromatin or "nonspecific" contamination. In this way, the ChAP-MS approach can help define and redefine mechanisms of chromatin-templated activities.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 4301 West Markham St, Little Rock, AR, 72205, USA.
authors.forename: Stephanie D
authors.initials: SD
authors.lastname: Byrum
authors.forename: Sean D
authors.initials: SD
authors.lastname: Taverna
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Tackett
mesh_terms: D018929:Cell Culture Techniques; D005591:Chemical Fractionation / Q000379:methods*; D002843:Chromatin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D005557:Formaldehyde / Q000494:pharmacology; D005686:Galactokinase / Q000235:genetics / Q000378:metabolism; D056426:Genetic Loci / Q000235:genetics; D059765:Homologous Recombination; D013058:Mass Spectrometry / Q000379:methods*; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D002843:Chromatin; D029701:Saccharomyces cerevisiae Proteins; D005557:Formaldehyde; C524219:GAL1 protein, S cerevisiae; D005686:Galactokinase