Isolation of Proteins on Nascent Chromatin and Characterization by Quantitative Mass Spectrometry

  • Paula A. Agudelo Garcia
  • Miranda Gardner
  • Michael A. Freitas
  • Mark R. ParthunEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1983)


Replication-coupled chromatin assembly is a very dynamic process that involves not only the replication fork machinery but also chromatin-related factors such as histones, histone chaperones, histone-modifying enzymes, and chromatin remodelers which ensure not only that the genetic information is properly replicated but also that the epigenetic code is reestablished in the daughter cell. Of the histone modifications associated with chromatin assembly, acetylation is the most abundant. Determining how newly synthesized histones get acetylated and what factors affect this modification is vital to understanding how cells manage to properly duplicate the epigenome. Here we describe a combination of the iPOND, quantitative mass spectrometry, and SILAC methodologies to study the protein composition of newly assembled chromatin and the modification state of the associated histones.


Nascent chromatin Acetylation SILAC Mass spectrometry Histone 



This work was supported by a grant from the National Institutes of Health (GM062970) to M.R.P.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Paula A. Agudelo Garcia
    • 1
  • Miranda Gardner
    • 2
  • Michael A. Freitas
    • 2
  • Mark R. Parthun
    • 1
    Email author
  1. 1.Department of Biological Chemistry and PharmacologyThe Ohio State UniversityColumbusUSA
  2. 2.Department of Cancer Biology and GeneticsThe Ohio State UniversityColumbusUSA

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