Abstract
The method for searching for ligands exerting an adjuvant effect is described. The method involves isolation of polysomes using an immobilized peptide mimetic of N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) — RN-peptide. After the affinity chromatography and washing, RN-peptide complexes with the target sequences were dissociated with guanidine hydrochloride. The obtained mRNA was used for cDNA synthesis and subsequent cloning in an expression vector. Further studies showed the effectiveness of this method. Clones interacting with the peptide were selected using biotinylated RN-peptide. It was found that all clones encode a sequence identical to the protein YB-1. Recombinant antibodies against protein YB-1 were selected from a phage display human scFv library. Using these antibodies, we determined the binding constant of RN-peptide to protein YB-1. Competitive analysis showed that RN-peptide and GMDP compete for the same portion of YB-1 at molar ratio 1: 12.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- GMDP:
-
N-acetylglucosaminyl-N-acetylmuramyl dipeptide
References
Audibert, F. (2003) Int. Immunopharmacol., 3, 1187–1193.
Cipriano, C., Giacconi, R., Muzzioli, M., Gasparini, N., Orlando, F., Corradi, A., Cabassi, E., and Mocchegiani, E. (2003) Mech. Ageing Dev., 124, 877–886.
Steiner, H. (2004) Immunol. Rev., 198, 83–96.
Golovina, T., Fattakhova, G., Swiderek, K., Makarov, E., Bovin, N., Shively, J., and Nesmeyanov, V. (1999) FEBS Lett., 454, 152–156.
Laman, A. G., Shepelyakovskaya, A. O., Boziyev, Kh. M., Savinov, G. V., and Brovko, F. A. (2010) Bioorg. Khim., 36, 141–148.
Monzavi-Karbassi, B., Cunto-Amesty, G., Luo, P., and Kieber-Emmons, T. (2002) Trends Biotechnol., 20, 207–214.
Palmiter, R. D. (1974) Biochemistry (Wash.), 13, 3606.
Laman, A. G., Shepelyakovskaya, A. O., Boziev, Kh. M., Savinov, G. V., Baidakova, L. K., Chulin, A. N., Chulina, I. A., Korpela, T., Nesmeyanov, V. A., and Brovko, F. A. (2011) Vaccine, 29, 7779–7784.
Sullivan, K. E., and Peterlin, B. M. (1987) Mol. Cell Biol., 7, 3315–3319.
Dorn, A., Durand, B., Marfing, C., Lemeur, M., Benoist, C., and Mathis, D. (1987) Proc. Natl. Acad. Sci. USA, 84, 6249–6253.
Boss, J. M., and Strominger, J. L. (1986) Proc. Natl. Acad. Sci. USA, 83, 9139–9143.
Sherman, P. A., Basta, P. V., and Ting, J. P.-Y. (1987) Proc. Natl. Acad. Sci. USA, 84, 4254–4258.
Miwa, K., Doyle, C., and Strominger, J. L. (1987) Proc. Natl. Acad. Sci. USA, 84, 4939–4943.
Eliseeva, I. A., Kim, E. R., Guryanov, S. G., Ovchinnikov, L. P., and Lyabin, D. N. (2011) Biochemistry (Moscow), 76, 1402–1433.
Batanova, T. A., Ulitin, A. B., Morozova, V. V., Laman, A. G., Zhirakovskaia, E. V., Dubrovskaia, V. V., Brovko, F. A., and Tikunova, N. V. (2006) Mol. Gen. Mikrobiol. Virusol., 3, 35–41.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © G. V. Savinov, A. O. Shepelyakovskaya, Kh. M. Boziev, F. A. Brovko, A. G. Laman, 2014, published in Biokhimiya, 2014, Vol. 79, No. 2, pp. 178–186.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM13–283, January 12, 2014.
Rights and permissions
About this article
Cite this article
Savinov, G.V., Shepelyakovskaya, A.O., Boziev, K.M. et al. Search for ligand of N-acetylglucosaminyl-N-acetylmuramyl dipeptide using its peptide mimetic. Biochemistry Moscow 79, 131–138 (2014). https://doi.org/10.1134/S0006297914020060
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0006297914020060