Proteome Characterization of a Chromatin Locus Using the Proteomics of Isolated Chromatin Segments Approach

Methods Mol Biol. 2017:1550:19-33. doi: 10.1007/978-1-4939-6747-6_3.

Abstract

The biological functions of given genomic regions are ruled by the local chromatin composition. The Proteomics of Isolated Chromatin segments approach (PICh) is a powerful and unbiased method to analyze the composition of chosen chromatin segments, provided they are abundant (repeated) or that the organism studied has a small genome. PICh can be used to identify novel and unexpected regulatory factors, or when combined with quantitative mass spectrometric approaches, to characterize the function of a defined factor at the chosen locus, by quantifying composition changes at the locus upon removal/addition of that factor.

Keywords: Chromatin; Heterochromatin; Protein Purification; Proteomics; Repeated Sequences; Telomeres.

MeSH terms

  • Cell Culture Techniques
  • Cells, Cultured
  • Chromatin / genetics*
  • Chromatin / isolation & purification
  • Chromatin / metabolism*
  • Chromatin Immunoprecipitation / methods
  • Heterochromatin / genetics
  • Heterochromatin / metabolism
  • Humans
  • Mass Spectrometry
  • Nucleic Acid Hybridization / methods
  • Proteins / chemistry*
  • Proteins / isolation & purification*
  • Proteome*
  • Proteomics / methods*
  • Repetitive Sequences, Nucleic Acid
  • Telomere
  • Workflow

Substances

  • Chromatin
  • Heterochromatin
  • Proteins
  • Proteome