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Aggregation of Influenza A Virus Nuclear Export Protein

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Abstract

Influenza A virus nuclear export protein (NEP) plays an important role in the viral life cycle. Recombinant NEP proteins containing (His)6-tag at either N-or C-terminus were obtained by heterologous expression in Escherichia coli cells and their high propensity for aggregation was demonstrated. Dynamic light scattering technique was used to study the kinetics and properties of NEP aggregation in solutions under different conditions (pH, ionic strength, presence of low-molecular-weight additives and organic solvents). Using atomic force microscopy, the predominance of spherical aggregates in all examined NEP preparations was shown, with some amyloid-like structures being observed in the case of NEP-C protein. A number of structure prediction programs were used to identify aggregation-prone regions in the NEP structure. All-atom molecular dynamics simulations indicate a high rate of NEP molecule aggregation and reveal the regions preferentially involved in the intermolecular contacts that are located at the edges of the rod-like protein molecule. Our results suggest that NEP aggregation is determined by different types of interactions and represents an intrinsic property of the protein that appears to be necessary for its functioning in vivo.

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Abbreviations

a.a.:

amino acid

AFM:

atomic force microscopy

CHAPS:

3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate

DLS:

dynamic light scattering

GuHCl:

guanidine hydrochloride

NEP:

nuclear export protein

NEP-C and NEP-N:

recombinant NEP proteins with (His)6-tag at the C-and N-termini, respectively

PdI:

polydis-persity index

Rh :

hydrodynamic radius

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Author information

Authors and Affiliations

  1. Department of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119991, Moscow, Russia

    A. O. Golovko

  2. Department of Chemistry, Lomonosov Moscow State University, 119991, Moscow, Russia

    O. N. Koroleva

  3. Department of Physics, Lomonosov Moscow State University, 119991, Moscow, Russia

    A. P. Tolstova & E. V. Dubrovin

  4. Department of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    N. V. Kuz’mina

  5. Frumkin Institute of Physical Chemistry and Electrochemistry, Russian Academy of Sciences, 119071, Moscow, Russia

    N. V. Kuz’mina

  6. Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    V. L. Drutsa

Authors
  1. A. O. Golovko
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  2. O. N. Koroleva
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  3. A. P. Tolstova
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  4. N. V. Kuz’mina
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  5. E. V. Dubrovin
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  6. V. L. Drutsa
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Corresponding author

Correspondence to O. N. Koroleva.

Additional information

Original Russian Text © A. O. Golovko, O. N. Koroleva, A. P. Tolstova, N. V. Kuz’mina, E. V. Dubrovin, V. L. Drutsa, 2018, published in Biokhimiya, 2018, Vol. 83, No. 11, pp. 1746–1758.

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Golovko, A.O., Koroleva, O.N., Tolstova, A.P. et al. Aggregation of Influenza A Virus Nuclear Export Protein. Biochemistry Moscow 83, 1411–1421 (2018). https://doi.org/10.1134/S0006297918110111

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  • DOI: https://doi.org/10.1134/S0006297918110111

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