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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A. S. Karyagina^1,2,3, I. S. Boksha^1,4, T. M. Grunina¹, A. V. Demidenko¹, M. S. Poponova¹, O. V. Sergienko^1,3, A. M. Lyashchuk¹, Z. M. Galushkina¹, L. A. Soboleva¹, E. O. Osidak⁵, M. S. Bartov¹, A. V. Gromov^1*, and V. G. Lunin^1,3

¹Gamaleya Center of Epidemiology and Microbiology, Ministry of Health of the Russian Federation, 123098 Moscow, Russia; E-mail: alexander.v.gromov@gmail.com

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

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

⁴Mental Health Research Center, 115522 Moscow, Russia

⁵IMTEK Ltd., 121552 Moscow, Russia

^* To whom correspondence should be addressed.

Received August 5, 2016; Revision received December 6, 2016

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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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KEY WORDS: recombinant bone morphogenetic protein-2, heterologous expression, Escherichia coli

DOI: 10.1134/S0006297917050091

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