Small-angle X-Ray analysis of macromolecular structure: the structure of protein NS2 (NEP) in solution
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CTURE OF MACROMOLECULAR COMPOUNDS
Small-Angle X-Ray Analysis of Macromolecular Structure: the Structure of Protein NS2 (NEP) in Solution E. V. Shtykovaa,b,*, E. N. Bogachevab, L. A. Dadinovaa, C. M. Jeffriesc, N. V. Fedorovad, A. O. Golovkoe, L. A. Baratovad, and O. V. Batishchevf a Shubnikov
Institute of Crystallography, Federal Scientific Research Centre “Crystallography and Photonics,” Russian Academy of Sciences, Moscow, 119333 Russia b Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, 119991 Russia c EMBL, Hamburg Outstation, c/o DESY, Hamburg, Germany d Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia e Faculty of Bioengineering and Bioinformatics, Moscow State University, Moscow, Russia f Frumkin Institute of Electrochemistry, Russian Academy of Sciences, Moscow, 119071 Russia * e-mail: [email protected] Received June 28, 2017
Abstract—A complex structural analysis of nuclear export protein NS2 (NEP) of influenza virus A has been performed using bioinformatics predictive methods and small-angle X-ray scattering data. The behavior of NEP molecules in a solution (their aggregation, oligomerization, and dissociation, depending on the buffer composition) has been investigated. It was shown that stable associates are formed even in a conventional aqueous salt solution at physiological рН value. For the first time we have managed to get NEP dimers in solution, to analyze their structure, and to compare the models obtained using the method of the molecular tectonics with the spatial protein structure predicted by us using the bioinformatics methods. The results of the study provide a new insight into the structural features of nuclear export protein NS2 (NEP) of the influenza virus A, which is very important for viral infection development. DOI: 10.1134/S1063774517060220
INTRODUCTION The influenza virus A (Orthomyxoviridae family) is a widespread pathogen, which manifests itself in periodically repeating lethal epidemics and pandemics. Despite the fact that the influenza virus has been studied for a long time, many stages of its life cycle, as well as the viral proteome components, are still insufficiently well known. In particular, the nuclear export protein NS2 (NEP) of influenza virus A (14 kDa, 121 amino acids), which is very important for the viral infection development, has been rather poorly investigated. This protein is known to participate in the viral genome export from nuclei of infected cells, playing the role of a mediator between the viral RNP (ribonucleoprotein composed of eight RNA fragments, bound with proteins NP, РВ1, РВ2, and РА) and cellular nuclear export proteins [1, 2]. However, the function of protein NS2 is not only to participate in export; this small protein contains interaction sites with at least five viral and cellular proteins: PB1, PB2, M1, CRM1, and F1Fo-ATPase [3]. These interaction sites are located both in the structured part of protein (C-terminal domain) and in its disordered part (N-terminal domain). T
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