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. 2021 Apr 19;2(2):100479.
doi: 10.1016/j.xpro.2021.100479. eCollection 2021 Jun 18.

Structural and biophysical characterization of the nucleosome-binding PZP domain

Brianna J Klein  1 , Khan L Cox  2 , Suk Min Jang  3 , Rohit K Singh  1 , Jacques Côté  3 , Michael G Poirier  2 , Tatiana G Kutateladze  1
Affiliations

Structural and biophysical characterization of the nucleosome-binding PZP domain

Brianna J Klein et al. STAR Protoc. .

Abstract

The core subunit of the MORF acetyltransferase complex BRPF1 contains a unique combination of zinc fingers, including a plant homeodomain (PHD) finger followed by a zinc knuckle and another PHD finger, which together form a PZP domain (BRPF1PZP). BRPF1PZP has been shown to bind to the nucleosome and make contacts with both histone H3 tail and DNA. Here, we describe biophysical and structural methods for characterization of the interactions between BRPF1PZP, H3 tail, DNA, and the intact nucleosome. For complete details on the use and execution of this protocol, please refer to Klein et al. (2020).

Keywords: Microscopy; NMR; Protein Biochemistry; Protein expression and purification; Structural Biology; X-ray Crystallography.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
1H,15N HSQC spectrum of the apo-state of BRPF1PZP Good dispersion of resonances indicates that the protein is folded.
Figure 2
Figure 2
EMSA with NCP207 incubated with increasing amounts of wild-type BRPF1PZP
Figure 3
Figure 3
Binding curve obtained for the interaction of wild-type BRPF1PZP with NCP207
Figure 4
Figure 4
HAT activity of the native wild-type and mutant MORF complexes from K562 cells on histones Liquid HAT assays, in which reactions were spotted on P81 filters and counted by scintillation as counts per minute (CPM).
See this image and copyright information in PMC

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