Vladivostok, Vladivostok, Russian Federation
Vladivostok, Vladivostok, Russian Federation
Protein Skp Yersinia pseudotuberculosis serves as the periplasmic chaperone in bacteria and has the ability to bind human and rabbit IgG. This work is aimed at studying the effect of this chaperone on the aggregation of human IgG at various pH values of the medium. To solve this problem, the aggregation kinetics of rSkp, IgG, and IgG in the presence of a chaperone in acidic, near neutral, and basic solutions was studied using the dynamic light scattering method. It was shown that in the presence of rSkp, the rate of IgG self-association and aggregation decreases significantly. The obtained results demonstrate the pH-dependent character of the chaperone and immunoglobulin-binding activities of rSkp. The most stable low-molecular complexes (RH up to 10 nm) between chaperone and human IgG are formed at acidic pH values of the medium. In the case of alkalinization of the reaction medium, the chaperone activity of rSkp decreases, and the aggregation of IgG although slows, but to a lesser extent than at acidic pH. The information obtained may be of interest to developers of stable and high-quality biopharmaceuticals based on immunoglobulins.
chaperone Skp, Yersinia pseudotuberculosis, aggregation of proteins, protein-protein interactions, dynamic light scattering
1. Harms N., Koningstein G., Dontje W., Muller M., Oudega B., Luirink J., de Cock H. The early interaction of the outer membrane protein Pho E with the periplasmic chaperone Skp occurs at cytoplasmic membrane. J. Biol. Chem., 2001, vol. 276, pp. 18804-18811.
2. Schafer U., Beck K., Muller M. Skp, a molecular chaperone of gram-negative bacteria, is required for the formation of soluble periplasmic intermediates of outer membrane proteins. J. Biol. Chem., 1999, vol. 274, pp. 24567-24574.
3. Colov'eva T.F., Novikova O.D., Portnyagina O.Yu. Biogenez β-barrel'nyh integral'nyh belkov naruzhnoy membrany bakteriy. Biohimiya, 2012, t. 77, № 11, s. 1459-1477. [Solov’eva T.F., Novikova O.D., Portmyagina O.Yu. Biogenesis of β-Barrel Integral Proteins Bacterial Outer Membrane. Biochemistry (Moscow), 2012, vol. 77, no. 11, pp. 1459-1477. (In Russ.)]
4. Geyer R., Galanos C., Westphal O., Golecki J. A lipopolysaccharide-binding cell-surface protein from Salmonella minnesota. Isolation, partial characterization and occurrence in different Enterobacteriaceae. Eur. J. Biochem., 1979, vol. 98, pp. 27-38.
5. Holck A., Lossius I., Aasland R., Kleppe K. Purification and characterization of the 17 K protein, a DNA-binding protein from Escherichia coli. Biochim. Biophys. Acta, 1987, vol. 914, pp. 49-54.
6. Holck A., Lossius I., Aasland R., Haarr L., Kleppe K. DNA- and RNA-binding proteins of chromatin from Escherichia coli. Biochim. Biophys. Acta, 1987, vol. 908, pp. 188-199.
7. Holck A, Kleppe K. Identity of the 17-kilodalton protein, a DNA-binding protein from Escherichia coli, and the firA gene product. Gene, 1988, vol. 67, pp. 117-124.
8. Shrestha A., Shi L., Tanase S., Tsukamoto M., Nishino N., Tokita K., Yamamoto T. Bacterial Chaperone Protein, Skp, Induces Leukocyte Chemotaxis via C5a Receptor. Am. J. Pathol., 2004, vol. 164, pp. 763-772.
9. Sidorin E.V., Ziganshin R.H., Naberezhnyh G.A., Lihackaya G.N., Trifonov E.V., Anastyuk S.D., Chernikov O.V., Solov'eva T.F. Belok shaperon Skp iz Yersinia pseudotuberculosis obladaet sposobnost'yu svyazyvat' immunoglobuliny G. Biohimiya, 2009, t. 74, № 4, s. 501-514. [Sidorin E.V., Ziganshin R.Kh., Naberezhnyk G.A., Likhatskaya G.N., Trifonov E.V., Anastyuk S.D., Chernikov O.V., Solovyeva T.F. Chaperone protein Skp from Yersinia pseudotuberculosis is able to bind immunoglobulins G. Biochemistry (Moscow), 2009, vol. 74, no. 4, pp. 501-514. (In Russ.)]
10. Sidorin E.V., Tischenko N.M., Homenko V.A., Isaeva M.P., Dmitrenok P.S., Kim N.Yu., Lihackaya G.N., Solov'eva T.F. Molekulyarnoe klonirovanie, vydelenie i harakteristika shaperona Skp iz Yersinia pseudotuberculosis. Biohimiya, 2012, t. 77, № 11, s. 1571-1583. [Sidorin E.V., Tishchenko N.M., Khomenko V.A., Isayeva M.P., Dmitrenok P.S., Kim N.Yu., Likhatskaya G.N., Solovyeva T.F. Molecular cloning, isolation and characterization of chaperone Skp from Yersinia pseudotuberculosis. Biochemistry (Moscow), 2012, vol. 77, no. 11, pp. 1571-1583. (In Russ.)]
11. Hawea A., Kasperb J.C., Friessb W., Jiskoot W. Structural properties of monoclonal antibody aggregates induced by freeze-thawing and thermal stress. European Journal of Pharmaceutical Sciences, 2009, vol. 38, pp. 79-87.
12. Arosio P., Barolo G., Muller-Spath T., Wu H., Morbidelli M. Aggregation Stability of a Monoclonal Antibody During Downstream Processing. Pharm. Res., 2011, vol.28, pp. 1884-1894.
13. Esfandiary R., Parupudi A., Casas-Finet J., Gadre D., Sathish H. Mechanism of Reversible Self-Association of a Monoclonal Antibody: Role of Electrostatic and Hydrophobic Interactions. J. Pharm. Sci., 2015, vol. 104, pp. 577-586.
14. Poon S., Rybchyn M.S., Easterbrook-Smith S.B., Carver J.A., Pankhurst G.J., Wilson M.R. Mildly acidic pH activates the extracellular molecular chaperone clusterin, J. Biol. Chem., 2002, vol. 277, no. 42, pp. 39532-39540.
15. Tapley T.L., Franzmann T.M., Chakraborty S., Jakob U., Bardwell J.C.A. Protein refolding by pH-triggered chaperone binding and release. PNAS, 2010, vol. 107, no. 3, pp. 1071-1076.
16. Bose D., Patra M., Chakrabarti A. Effect of pH on stability, conformation, and chaperone activity of erythroid & non-erythroid spectrin. Biochim. Biophys. Acta - Proteins and Proteomics, 2017, vol. 1865, pp. 694-702.
