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Biochemistry (Moscow)

, Volume 82, Issue 5, pp 613–624 | Cite as

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

  • 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. GromovEmail author
  • V. G. Lunin
Article

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.

Keywords

recombinant bone morphogenetic protein-2 heterologous expression Escherichia coli 

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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Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • A. S. Karyagina
    • 1
    • 2
    • 3
  • I. S. Boksha
    • 1
    • 4
  • T. M. Grunina
    • 1
  • A. V. Demidenko
    • 1
  • M. S. Poponova
    • 1
  • O. V. Sergienko
    • 1
    • 3
  • A. M. Lyashchuk
    • 1
  • Z. M. Galushkina
    • 1
  • L. A. Soboleva
    • 1
  • E. O. Osidak
    • 5
  • M. S. Bartov
    • 1
  • A. V. Gromov
    • 1
    Email author
  • V. G. Lunin
    • 1
    • 3
  1. 1.Gamaleya Center of Epidemiology and MicrobiologyMinistry of Health of the Russian FederationMoscowRussia
  2. 2.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia
  3. 3.Russian Research Institute of Agricultural BiotechnologyMoscowRussia
  4. 4.Mental Health Research CenterMoscowRussia
  5. 5.IMTEK Ltd.MoscowRussia

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