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Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones
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Chapters
- 00:05Title
- 01:03Reconstitution of a Histone Octamer with Differentially Isotope-labeled and Asymmetrically-modified Histone H3
- 03:20Tandem Affinity Purification of Reconstituted Octamer
- 05:28Nucleosome Reconstitution
- 07:07Results: Generation of Differentially Isotope-labeled Nucleosomes for Investigating Cross-talk Mechanisms
- 08:58Conclusion
This protocol describes the reconstitution of nucleosomes containing differentially isotope-labeled sister histones. At the same time, asymmetrically post-translationally modified nucleosomes can be generated after using a premodified histone copy. These preparations can be readily used to study modification crosstalk mechanisms, simultaneously on both sister histones, by using high-resolution NMR spectroscopy.
