Izvorni znanstveni članak

https://doi.org/10.5562/cca2806

Simplified Method for Rapid Purification of Soluble Histones

Nives Ivić ; Gene Center Munich and Department of Biochemistry, Ludwig-Maximilians-Universität München, 81377 Munich, Germany
Bernhard Groschup ; Gene Center Munich and Department of Biochemistry, Ludwig-Maximilians-Universität München, 81377 Munich, Germany
Silvija Bilokapić orcid.org/0000-0003-2199-2929 ; Gene Center Munich and Department of Biochemistry, Ludwig-Maximilians-Universität München, 81377 Munich, Germany
Mario Halić ; Gene Center Munich and Department of Biochemistry, Ludwig-Maximilians-Universität München, 81377 Munich, Germany

Sažetak

Functional and structural studies of histone-chaperone complexes, nucleosome modifications, their interactions with remodelers and regulatory proteins rely on obtaining recombinant histones from bacteria. In the present study, we show that co-expression of Xenopus laevis histone pairs leads to production of soluble H2AH2B heterodimer and (H3H4)2 heterotetramer. The soluble histone complexes are purified by simple chromatographic techniques. Obtained H2AH2B dimer and H3H4 tetramer are proficient in histone chaperone binding and histone octamer and nucleosome formation. Our optimized protocol enables rapid purification of multiple soluble histone variants with a remarkable high yield and simplifies histone octamer preparation. We expect that this simple approach will contribute to the histone chaperone and chromatin research.

This work is licensed under a Creative Commons Attribution 4.0 International License.

Ključne riječi

histone; nucleosome; chromatin; protein co-expression

Hrčak ID:

159897

URI

https://hrcak.srce.hr/159897

Datum izdavanja:

21.6.2016.

Posjeta: 5.253 *