Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 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.

PubMed Disclaimer

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

Similar articles

See all similar articles

Cited by

References

    1. Adams P.D., Afonine P.V., Bunkoczi G., Chen V.B., Davis I.W., Echols N., Headd J.J., Hung L.W., Kapral G.J., Grosse-Kunstleve R.W. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 2010;66:213–221. - PMC - PubMed
    1. Andrews F.H., Strahl B.D., Kutateladze T.G. Insights into newly discovered marks and readers of epigenetic information. Nat. Chem. Biol. 2016;12:662–668. - PMC - PubMed
    1. Avvakumov N., Lalonde M.E., Saksouk N., Paquet E., Glass K.C., Landry A.J., Doyon Y., Cayrou C., Robitaille G.A., Richard D.E. Conserved molecular interactions within the HBO1 acetyltransferase complexes regulate cell proliferation. Mol. Cell. Biol. 2012;32:689–703. - PMC - PubMed
    1. Behal R.H., Betleja E., Cole D.G. Purification of IFT particle proteins and preparation of recombinant proteins for structural and functional analysis. Methods Cell Biol. 2009;93:179–196. - PubMed
    1. Berman H., Henrick K., Nakamura H. Announcing the worldwide Protein Data Bank. Nat. Struct. Biol. 2003;10:980. - PubMed

Publication types

MeSH terms

LinkOut - more resources