Purification of protein–DNA complexes by native gel electrophoresis for electron microscopy study

被引：0

作者：

Valieva M.E. ^{[1
]}

Derkacheva N.I. ^{[2
]}

Sokolova O.S. ^{[1
]}

机构：

[1] Department of Biology, Moscow State University, Moscow

[2] Department of Biochemistry, Evdokimov University of Medicine and Dentistry, Moscow

来源：

Moscow University Biological Sciences Bulletin | 2017年 / 72卷 / 1期

关键词：

dinucleosome; electron microscopy; image processing; mononucleosome; PAGE;

D O I：

10.3103/S0096392517010059

中图分类号：

学科分类号：

摘要：

Electrophoretic separation under native conditions may be used for purification of protein molecules and their complexes with DNA and other ligands. Here, we employed this approach to separate protein-DNA complexes with a molecular weight of approximately 200 kDa: mono- and dinucleosomes. The purified mononucleosomes were subjected to single particle electron microscopy study using negative stain contrasting, and the two-dimensional projections of the nucleosomes at 25 Å resolution were obtained. A comparison of the nucleosome projections before and after separation in the native PAGE revealed different orientation of particles on the carbon film. © 2017, Allerton Press, Inc.

引用

页码：1 / 5

页数：4

共 19 条

[1]

Clapier C., Cairns B., The biology of chromatin remodeling complexes, Annu. Rev. Biochem., 78, pp. 273-304, (2009)

[2]

Vignali M., Hassan A., Neely K., Workman J., ATP-dependent chromatin-remodeling complexes, Mol. Cell. Biol., 20, 6, pp. 1899-1910, (2000)

[3]

Kornberg R.D., Thomas J.O., Chromatin structure: Oligomers of the histones, Science, 184, 4139, pp. 865-868, (1974)

[4]

Luger K., Mader A.W., Richmond R.K., Sargent D.F., Richmond T.J., Crystal structure of the nucleosome core particle at 2.8 Å resolution, Nature, 389, 6648, pp. 251-260, (1997)

[5]

Arents G., Moudrianakis E.N., The histone fold: A ubiquitous architectural motif utilized in DNA compaction and protein dimerization, Proc. Natl. Acad. Sci. U.S.A., 92, 24, pp. 11170-11174, (1995)

[6]

Dechassa M.L., Zhang B., Horowitz-Scherer R., Persinger J., Woodcock C.L., Peterson C.L., Bartholomew B., Architecture of the SWI/SNFnucleosome complex, Mol. Cell. Biol., 28, 19, pp. 6010-6021, (2008)

[7]

Feser J., Truong D., Das C., Carson J.J., Kieft J., Harkness T., Tyler J.K., Elevated histone expression promotes life span extension, Mol. Cell, 39, 5, pp. 724-735, (2010)

[8]

Ljungman M., Hanawalt P., Efficient protection against oxidative DNA damage in chromatin, Mol. Carcinog., 5, 4, pp. 264-269, (1992)

[9]

Enright H.U., Miller W.J., Hebbel R.P., Nucleosomal histone protein protects DNA from iron mediated damage, Nucleic Acids Res., 20, 13, pp. 3341-3346, (1992)

[10]

van Heel M., Gowen B., Matadeen R., Orlova E.V., Finn R., Pape T., Cohen D., Stark H., Schmidt R., Schatz M., Patwardhan A., Single-particle electron cryo-microscopy: Towards atomic resolution, Q. Rev. Biophys., 33, 4, pp. 307-369, (2000)

← 1 2 →