Abstract
Methods for preparation of recombinant physiologically active proteins of the Noggin family (Noggin1 and Noggin2) of the clawed frog Xenopus laevis that are capable of interaction with BMP factors of the TGF-beta superfamily have been developed. Structures were designed for the expression of Noggin1 and Noggin2 proteins allowing us to produce them from the directly microinjected synthetic mRNA in the cells of developing embryos of the clawed frog, as well as in the prokaryotic expression system. Target proteins contained three Myc-epitopes at the N-terminal. Introduction of these’ tags’ allowed comparison of the expression level of the Noggin1 and Noggin2 proteins, isolation on the affinity immunosorbent, and demonstration of biological activity of the isolated Noggin-proteins through the analysis of their ability to bind the BMP4 factor of the TGF-beta superfamily with coimmunoprecipitation method.
Similar content being viewed by others
Abbreviations
- BMP:
-
bone morphogenic protein
- TGF:
-
transforming growth factor
- Xnr2:
-
xenopus nodal-related
References
Eroshkin, F.M., Bayramov, A.V., Martynova, N.Y., and Zaraisky, A.G., Russ. J. Bioorg. Chem., 2012, vol. 38, pp. 338–340.
Bayramov, A.V., Eroshkin, F.M., Martynova, N.Y., Ermakova, G.V., Solovieva, E.A., and Zaraisky, A.G., Development, 2011, vol. 138, no. 24, pp. 5345–5356.
Watanabe, K., Kamiya, D., Nishiyama, A., Katayama, T., Nozaki, S., Kawasaki, H., Watanabe, Y., Mizuseki, K., and Sasai, Y., Nature Neuroscience, 2005, vol. 8, pp. 288–296.
Dreesen, O. and Brivanlow, A.H., Stem Cell Rev., 2007, vol. 3, pp. 7–17.
Smith, W.C. and Harland, R.M., Cell, 1992, vol. 70, pp. 829–840.
Martynova, N.Y., Eroshkin, F.M., Ermolina, L.V., Ermakova, G.V., Korotaeva, A.L., Smurova, K.M., Gyoeva, F.K., and Zaraisky, A.G., Dev. Dynamics, 2008, vol. 237, pp. 736–749.
Matthews, G.M. and Colman, A., Method Mol. Biol., 1998, vol. 86, pp. 235–247.
Groppe, J., Greenwald, J., Wiater, E., Rodriguez-Leon, J., and Economides, A.N., Nature, 2002, vol. 420, pp. 636–642.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © F.M. Eroshkin, A.V. Bairamov, O.V. Aver’yanova, E.A. Solov’eva, M.V. Serebryakova, A.G. Zaraiskii, N.Yu. Martynova, 2013, published in Bioorganicheskaya Khimiya, 2013, Vol. 39, No. 2, pp. 247–252.
Rights and permissions
About this article
Cite this article
Eroshkin, F.M., Bairamov, A.V., Aver’yanova, O.V. et al. Preparation and analysis of biological activity of the secreted proteins of the Noggin family. Russ J Bioorg Chem 39, 218–222 (2013). https://doi.org/10.1134/S1068162013020040
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1068162013020040