Abstract
Control of gene expression and the faithful transmission of genetic and epigenetic information rely on chromatin-bound proteins. These include the polycomb group of proteins, which can display a remarkable variability in their composition. Alterations in the chromatin-bound protein compositions are relevant for physiology and human disease. Thus, chromatin-bound proteomic profiling can be instrumental for understanding fundamental cellular processes and for identifying therapeutic targets. Inspired by biochemical strategies for the isolation of proteins on nascent DNA (iPOND) and the very similar DNA-mediated chromatin pull-down (Dm-ChP), we described a method for the identification of Protein on Total DNA (iPOTD) for bulk chromatome profiling. Here, we update our iPOTD method and, in particular, detail the experimental procedure for the isolation of chromatin proteins for mass spectrometry-based proteomic analysis.
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Acknowledgments
This work was supported by the Spanish of Economy, Industry and Competitiveness (MEIC) (PID2019-108322GB-100), “CaixaResearch Health” (HR20-00411), “Fundación Vencer El Cancer” (VEC), the European Regional Development Fund (FEDER), and from AGAUR to L.D.C. The Ramon y Cajal program of the Ministerio de Ciencia, Innovación y Universidades and the European Social Fund under the reference number RYC-2018-025002-I, and the Instituto de Salud Carlos III-FEDER (PI19/01814 and PI22/01837), to S.A. We acknowledge the funding support of the Spanish Ministry of Science and Innovation to the EMBL partnership, the Centro de Excelencia Severo Ochoa, and the CERCA Programme/Generalitat de Catalunya.
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Aranda, S., Di Croce, L. (2023). Isolation of Chromatin Proteins by Genome Capture. In: Lanzuolo, C., Marasca, F. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 2655. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3143-0_8
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DOI: https://doi.org/10.1007/978-1-0716-3143-0_8
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