Determining the Binding Constant of LANA Protein Fragment with Nucleosome

作     者：Novikov, R.V. Bondarenko, E.A. Malyuchenko, N.V. Feofanov, A.V. Studitsky, V.M. Shaytan, A.K. 

作者机构：Bioengineering Department Biological Faculty Moscow State University Moscow 119234 Russian Federation Sirius Science and Technology University Sochi 354340 Russian Federation Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences Moscow 117997 Russian Federation Cancer Epigenetics Program Fox Chase Cancer Center Philadelphia 19111 PA United States 

出 版 物：《Moscow University Biological Sciences Bulletin》 (Moscow Univ. Biol. Sci. Bull.)

年 卷 期：2020年第75卷第4期

页      面：252-256页

学科分类：0710[理学-生物学] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 07[理学] 09[农学] 

基　　金：Russian Foundation for Basic Research  РФФИ  (19-74-30003) 

主　　题：acidic patch binding constant gel electrophoresis Hill equation LANA nucleosome 

摘      要：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 an 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 therapeutic agents design. In this work, we propose a technique and measure the binding constant of the LANA protein fragment to the 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. © 2020, Allerton Press, Inc.