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