Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional protein domains: Synthesis in an Escherichia coli heterologous expression system
- Authors: Karyagina A.S.1,2,3, Boksha I.S.1,4, Grunina T.M.1, Demidenko A.V.1, Poponova M.S.1, Sergienko O.V.1,3, Lyashchuk A.M.1, Galushkina Z.M.1, Soboleva L.A.1, Osidak E.O.5, Bartov M.S.1, Gromov A.V.1, Lunin V.G.1,3
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Affiliations:
- Gamaleya Center of Epidemiology and Microbiology
- Belozersky Institute of Physico-Chemical Biology
- Russian Research Institute of Agricultural Biotechnology
- Mental Health Research Center
- IMTEK Ltd.
- Issue: Vol 82, No 5 (2017)
- Pages: 613-624
- Section: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151384
- DOI: https://doi.org/10.1134/S0006297917050091
- ID: 151384
Cite item
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.
About the authors
A. S. Karyagina
Gamaleya Center of Epidemiology and Microbiology; Belozersky Institute of Physico-Chemical Biology; Russian Research Institute of Agricultural Biotechnology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098; Moscow, 119992; Moscow, 127550
I. S. Boksha
Gamaleya Center of Epidemiology and Microbiology; Mental Health Research Center
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098; Moscow, 115522
T. M. Grunina
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
A. V. Demidenko
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
M. S. Poponova
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
O. V. Sergienko
Gamaleya Center of Epidemiology and Microbiology; Russian Research Institute of Agricultural Biotechnology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098; Moscow, 127550
A. M. Lyashchuk
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
Z. M. Galushkina
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
L. A. Soboleva
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
E. O. Osidak
IMTEK Ltd.
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 121552
M. S. Bartov
Gamaleya Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
A. V. Gromov
Gamaleya Center of Epidemiology and Microbiology
Author for correspondence.
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098
V. G. Lunin
Gamaleya Center of Epidemiology and Microbiology; Russian Research Institute of Agricultural Biotechnology
Email: alexander.v.gromov@gmail.com
Russian Federation, Moscow, 123098; Moscow, 127550