Biochemistry (Moscow)

, Volume 77, Issue 11, pp 1315–1325

Molecular cloning, isolation, and properties of chaperone Skp from Yersinia pseudotuberculosis

Authors

    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • N. M. Tishchenko
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • V. A. Khomenko
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • M. P. Isaeva
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • P. S. Dmitrenok
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • N. Yu. Kim
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • G. N. Likhatskaya
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
  • T. F. Solov’eva
    • Elyakov Pacific Institute of Bioorganic ChemistryFar Eastern Branch of the Russian Academy of Sciences
Article

DOI: 10.1134/S0006297912110119

Cite this article as:
Sidorin, E.V., Tishchenko, N.M., Khomenko, V.A. et al. Biochemistry Moscow (2012) 77: 1315. doi:10.1134/S0006297912110119

Abstract

The skp gene of Yersinia pseudotuberculosis was expressed without its signal sequence in Escherichia coli BL21(DE3) cells. The recombinant protein Skp accumulated in soluble form in the cytoplasm of the producer strain. The protein was isolated and characterized: the molecular weight, isoelectric point, N-terminal amino acid sequence (20 amino acid residues), and the content of the secondary structure elements were determined. Using cross-linking stabilization and high-mass MALDI-TOF mass spectrometric analysis, it was found that rSkp forms a stable homotrimer in solution and interacts with human IgG. Three-dimensional models of the Skp trimer and its complexes with Fc- and Fab-fragments of human IgG1 were constructed by computer modeling.

Key words

chaperone SkpYersinia pseudotuberculosisimmunoglobulin Gcross-linking stabilizationMALDI-TOF mass spectrometrycomputer modelingprotein-protein interactions

Abbreviations

IPTG

isopropyl-β-D-1-thiogalactopyranoside

MALDI-TOF MS

matrix-assisted laser desorption ionization time-of-flight mass spectrometry

rSkp

recombinant chaper-one Skp

Copyright information

© Pleiades Publishing, Ltd. 2012