A family of thermal stable Sm-like oligomeric proteins is an extremely attractive object for protein engineering. Earlier, we demonstrated the possibility of using them as a carrier for reducing the aggregation ability and increasing the thermal stability of amyloidogenic peptides. For further work, we need to find the structural basis for the stability and specificity of oligomerization of these proteins. To find the basis for specificity, we proposed a new approach. The first step is to find such pairs of Sm-like proteins that can form hetero-oligomers. In this paper, we describe the approach we have developed to achieve this goal. The obtained experimental result will allow us to proceed to the second step - the theoretical analysis of protein structures and the identification of structural features that determine the intersubunit interaction in the oligomer.
protein engineering, Sm-like proteins, protein folding, oligomerization, interaction specificity
1. Achsel T., Brahms H., Kastner B., Bachi A., Wilm M., Lührmann R.A doughnut-shaped heteromer of human Sm-like proteins binds to the 3￠-end of U6 snRNA, thereby facilitating U4/U6 duplex formation in vitro. EMBO J., 1999, vol. 18, pp. 5789-5802.
2. Murina V.N, Nikulin A.D. RNK-svyazyvayuschie Sm-podobnye belki bakteriy i arhey: shodstvo i razlichiya struktur i funkciy. Uspehi biologicheskoy himii, 2011, t. 51, s. 133-164. [Murina V.N, Nikulin A.D. RNA-binding bacterial and archaeous Sm-like proteins, commons and differs of structure and functions. Bio. Che. Succ., 2011, vol. 51, pp. 133-164. (In Russ.)]
3. Toro I., Basquin J., Teo-Dreher H., Suck D. Archaeal Sm proteins form heptameric and hexameric complexes: crystal structures of the Sm1 and Sm2 proteins from the hyperthermophile Archaeoglobus fulgidus. J. Mol. Biol., 2002, vol. 320, pp. 129-42.
4. Zhang A., Wassarman K.M., Ortega J., Steven A.C, Storz G. The Sm-like Hfq protein increases OxyS RNA interaction with target mRNAs. Molecular cell., 2002, vol. 9, pp. 11-22.
5. Seraphin B. Sm and Sm-like proteins belong to a large family: identification of proteins of the U6 as well as the U1, U2, U4 and U5 snRNPs. EMBO J., 1995, vol. 14, pp. 2089-2098.
6. Link T.M., Valentin-Hansen P., Brennan R.G. Structure of Escherichia coli Hfq bound to polyriboadenylate RNA. Proc. Natl. Acad. Sci., 2009, vol. 106, pp. 19292-19297.
7. Nikulin A., Stolboushkina E., Perederina A., Vassilieva I., Blaesi U., Moll I., Kachalova G., Yokoyama S., Vassylyev D., Garber M., Nikonov S. Structure of Pseudomonas aeruginosa Hfq protein. Acta Crystallogr D Biol Crystallogr., 2005, vol. 61, pp. 141-146.
8. Brennan R.G., Link T.M. Hfq structure, function and ligand binding. Curr Opin Microbiol., 2007, vol. 10, pp. 125-133.
9. Murina V.N, Mel'nik B.S., Filimonov V.V., Ulayn M., Veyss M.S., Myuller U., Nikulin A.D. Vliyanie zamen konservativnyh aminokislotnyh ostatkov na strukturu i stabil'nost' belka Hfq. Biohimiya, 2014, t. 79, vyp. 5, c. 595-604. [Murina V.N., Melnik B.S., Filimonov V.V., Ulane M., Vase M.S., Muller U., Nikulin A.D. Influence of conservative aminoacids changes on the structure and stability of Hfq protein. Biochemistry, 2014, vol. 79, iss. 5, pp.595-604. (In Russ.)]
10. Moskaleva O., Melnik B., Gabdulkhakov A., Garber M., Nikonov S., Stolboushkina E., Nikulin A. The structures of mutant forms of Hfq from Pseudomonas aeruginosa reveal the importance of the conserved His57 for the protein hexamer organization. Structural Biology and Crystallization Communication, 2010, vol. 66, rr. 760-764.
