Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes

R. K. McGinty, R. D. Makde, Song Tan

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

Abstract

Structural studies of chromatin complexes composed of chromatin factors or enzymes bound to the nucleosome have been constrained by the ability to produce high-quality complexes in the amounts appropriate for biophysical studies and by the difficulty of crystallizing these complexes. We describe here procedures and approaches to prepare chromatin complexes, to crystallize chromatin complexes, and to improve diffraction properties through postcrystallization soaks. Special attention is paid to evaluating the quality of the purified chromatin complexes as well as assessing the presence of the chromatin protein or enzyme in crystals. The methods described for preparing and purifying chromatin complexes should be applicable to biochemical, biophysical, and other structural approaches including cryoelectron microscopy.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages43-65
Number of pages23
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Enzymology
Volume573
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Nucleosomes
Crystallization
Chromatin
Enzymes
Cryoelectron Microscopy
Microscopic examination
Diffraction
Crystals

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

McGinty, R. K., Makde, R. D., & Tan, S. (2016). Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes. In Methods in Enzymology (pp. 43-65). (Methods in Enzymology; Vol. 573). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.01.003
McGinty, R. K. ; Makde, R. D. ; Tan, Song. / Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes. Methods in Enzymology. Academic Press Inc., 2016. pp. 43-65 (Methods in Enzymology).
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McGinty, RK, Makde, RD & Tan, S 2016, Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes. in Methods in Enzymology. Methods in Enzymology, vol. 573, Academic Press Inc., pp. 43-65. https://doi.org/10.1016/bs.mie.2016.01.003

Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes. / McGinty, R. K.; Makde, R. D.; Tan, Song.

Methods in Enzymology. Academic Press Inc., 2016. p. 43-65 (Methods in Enzymology; Vol. 573).

Research output: Chapter in Book/Report/Conference proceedingChapter

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AB - Structural studies of chromatin complexes composed of chromatin factors or enzymes bound to the nucleosome have been constrained by the ability to produce high-quality complexes in the amounts appropriate for biophysical studies and by the difficulty of crystallizing these complexes. We describe here procedures and approaches to prepare chromatin complexes, to crystallize chromatin complexes, and to improve diffraction properties through postcrystallization soaks. Special attention is paid to evaluating the quality of the purified chromatin complexes as well as assessing the presence of the chromatin protein or enzyme in crystals. The methods described for preparing and purifying chromatin complexes should be applicable to biochemical, biophysical, and other structural approaches including cryoelectron microscopy.

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McGinty RK, Makde RD, Tan S. Preparation, Crystallization, and Structure Determination of Chromatin Enzyme/Nucleosome Complexes. In Methods in Enzymology. Academic Press Inc. 2016. p. 43-65. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2016.01.003