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Chaperone and Immunoglobulin-Binding Activities of Skp Protein from Yersinia pseudotuberculosis

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Abstract

Here, we determined qualitative and quantitative characteristics of the chaperone and immunoglobulin-binding activities of recombinant Skp protein (rSkp) from Yersinia pseudotuberculosis using the methods of dynamic light scattering and surface plasmon resonance. Commercial human polyclonal IgG and Fc and Fab fragments of human IgG were used as substrate proteins. The activity of rSkp strongly depended on the medium pH. The most stable low-molecular-weight complexes with a hydrodynamic radius up to 10 nm were formed by rSkp and protein substrates at acidic pH values. Under these conditions, rSkp exhibited the lowest propensity to self-association and the highest affinity for human IgG and its Fc and Fab fragments, as well as prevented their aggregation most efficiently (i.e., demonstrated the maximal chaperone activity). As the medium pH increased, the affinity of rSkp for IgG and its fragments decreased; rSkp was not able to completely prevent the aggregation of protein substrates, but significantly slowed it down. The obtained information may be of practical interest, since the stability of therapeutic IgG preparations affects their safety and efficacy in medical applications.

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Abbreviations

DLS:

dynamic light scattering

IgG:

human immunoglobulin G

k a :

rate constant of complex association

K D :

kinetic constant of complex dissociation

k d :

rate constant of complex dissociation

RH:

hydrodynamic radius of particles (in distribution analysis)

rSkp:

recombinant Skp protein (17 kDa)

RU:

resonance unit

SPR:

surface plasmon resonance

Z-average:

average hydrodynamic radius of particles (in analysis of cumulants).

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Authors and Affiliations

  1. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, 690022, Vladivostok, Russia

    E. V. Sidorin, V. A. Khomenko, N. Yu. Kim & T. F. Solov’eva

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  1. E. V. Sidorin
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  2. V. A. Khomenko
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  3. N. Yu. Kim
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  4. T. F. Solov’eva
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Correspondence to E. V. Sidorin.

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Conflict of interest. The authors declare no conflict of interest.

Ethical norm compliance. This article does not contain description of studies with articipation of humans or animals performed by any of the authors.

Russian Text © The Author(s), 2020, published in Biokhimiya, 2020, Vol. 85, No. 1, pp. 93–103.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM19-150, December 9, 2019.

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Sidorin, E.V., Khomenko, V.A., Kim, N.Y. et al. Chaperone and Immunoglobulin-Binding Activities of Skp Protein from Yersinia pseudotuberculosis. Biochemistry Moscow 85, 80–89 (2020). https://doi.org/10.1134/S0006297920010071

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