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Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional protein domains: Synthesis in an Escherichia coli heterologous expression system

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Abstract

Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional N-terminal protein domains were obtained by expression in E. coli. The N-terminal domains were s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) and lz (leucine zipper dimerization domain from yeast transcription factor GCN4). The s-tag-BMP-2 and lz-BMP-2 were purified by a procedure that excluded a long refolding stage. The resulting dimeric proteins displayed higher solubility compared to rhBMP-2 without additional protein domains. Biological activity of both proteins was demonstrated in vitro by induction of alkaline phosphatase in C2C12 cells, and the activity of s-tag-BMP-2 in vivo was shown in various experimental animal models.

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Abbreviations

a.a.:

amino acid residue

BCA:

2,2′-bicinchoninic acid

BMP-2:

bone morphogenetic protein-2

hBMP-2:

human BMP-2

lz:

fragment of yeast GCN4 transcription factor corresponding to GCN4 leucine zipper dimerization domain

rhBMP-2:

recombinant human BMP-2

s-tag:

15-a.a. fragment of bovine pancreatic ribonuclease A

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Authors and Affiliations

  1. Gamaleya Center of Epidemiology and Microbiology, Ministry of Health of the Russian Federation, 123098, Moscow, Russia

    A. S. Karyagina, I. S. Boksha, T. M. Grunina, A. V. Demidenko, M. S. Poponova, O. V. Sergienko, A. M. Lyashchuk, Z. M. Galushkina, L. A. Soboleva, M. S. Bartov, A. V. Gromov & V. G. Lunin

  2. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992, Moscow, Russia

    A. S. Karyagina

  3. Russian Research Institute of Agricultural Biotechnology, 127550, Moscow, Russia

    A. S. Karyagina, O. V. Sergienko & V. G. Lunin

  4. Mental Health Research Center, 115522, Moscow, Russia

    I. S. Boksha

  5. IMTEK Ltd., 121552, Moscow, Russia

    E. O. Osidak

Authors
  1. A. S. Karyagina
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  2. I. S. Boksha
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  3. T. M. Grunina
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  4. A. V. Demidenko
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  5. M. S. Poponova
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  6. O. V. Sergienko
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  7. A. M. Lyashchuk
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  8. Z. M. Galushkina
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  9. L. A. Soboleva
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  10. E. O. Osidak
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  11. M. S. Bartov
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  12. A. V. Gromov
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  13. V. G. Lunin
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Corresponding author

Correspondence to A. V. Gromov.

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Original Russian Text © A. S. Karyagina, I. S. Boksha, T. M. Grunina, A. V. Demidenko, M. S. Poponova, O. V. Sergienko, A. M. Lyashchuk, Z. M. Galushkina, L. A. Soboleva, E. O. Osidak, M. S. Bartov, A. V. Gromov, V. G. Lunin, 2017, published in Biokhimiya, 2017, Vol. 82, No. 5, pp. 817-831.

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Karyagina, A.S., Boksha, I.S., Grunina, T.M. et al. Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional protein domains: Synthesis in an Escherichia coli heterologous expression system. Biochemistry Moscow 82, 613–624 (2017). https://doi.org/10.1134/S0006297917050091

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  • DOI: https://doi.org/10.1134/S0006297917050091

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