Determination of the binding constant of LANA protein fragment with nucleosome
Abstract
On the surface of the nucleosome, there are many regions for interaction with chromatin proteins, but an acidic patch, a negatively charged region formed by the residues of the histone H2A/H2B dimer, is especially prominent. The acidic patch is a target for anticancer drugs and interaction locus for various pathogens. For instance, it was shown that this region is the binding site of the LANA protein of human gammaherpesvirus 8 and tethers the virus episomes to mitotic chromosomes. The development of methods for analyzing the binding of various compounds to nucleosomes is necessary both for understanding the mechanisms of chromatin regulation and for the therapeutic agents design. In this work, we propose a technique and measure the binding constant of the LANA protein fragment to nucleosome. In contrast to previous studies, the measurements were carried out at a physiological salt concentration in the buffer solution. The proposed technique is based on the signal detection from the fluorescently labeled peptides after the separation of complexes of nucleosomes with peptides by gel electrophoresis. The paper also discusses mathematical models for analyzing the interaction between peptides and nucleosome and possible factors that can affect it.
About the Authors
R. V. Novikov
Bioengineering Department, School of Biology, Lomonosov Moscow State University; “Sirius” Science and Technology University
Russian Federation
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Olympic str., 1, Sochi, 354340
E. A. Bondarenko
Bioengineering Department, School of Biology, Lomonosov Moscow State University
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
N. V. Malyuchenko
Bioengineering Department, School of Biology, Lomonosov Moscow State University
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
A. V. Feofanov
Bioengineering Department, School of Biology, Lomonosov Moscow State University; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Miklukho-Maklaya ul. 16/10, Moscow, 117997
V. M. Studitsky
Bioengineering Department, School of Biology, Lomonosov Moscow State University; Cancer Epigenetics Program, Fox Chase Cancer Center
Russian Federation
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Cottman Avenue 333, Philadelphia, PA 19111, USA
A. K. Shaytan
Bioengineering Department, School of Biology, Lomonosov Moscow State University; “Sirius” Science and Technology University
Russian Federation
Russian Federation
Leninskiye Gory 1–12, Moscow, 119234
Olympic str., 1, Sochi, 354340
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Review
For citations:
Novikov R.V., Bondarenko E.A., Malyuchenko N.V., Feofanov A.V., Studitsky V.M., Shaytan A.K. Determination of the binding constant of LANA protein fragment with nucleosome. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2020;75(4):296-301. (In Russ.)
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