OmpC-like porin from Yersinia pseudotuberculosis: Molecular characteristics, physico-chemical and functional properties

  • O. D. Novikova
  • V. A. Khomenko
  • V. I. Emelyanenko
  • G. N. Likhatskaya
  • E. A. Zelepuga
  • N. Yu. Kim
  • M. P. Isaeva
  • O. Yu. Portnyagina
  • O. P. Vostrikova
  • O. V. Sidorova
  • T. F. Solov’eva


Pore-forming protein from the outer membrane of Yersinia pseudotuberculosis cultured at 37°C has been isolated and characterized. Comparative analysis of the primary and three-dimensional structures of this protein and of OmpC porin from E. coli was carried out, functional properties of these proteins have been studied using bilayer lipid membranes (BLM) technique. The degree of homology, molecular mass and pore-forming properties of the isolated porin was found to be closer to those of OmpC porin from E. coli than OmpF porin from Y. pseudotuberculosis. The value of the most probable conductivity of OmpC porin from Y. pseudotuberculosis (0.18 pS) in BLM corresponded to the conductivity of the native trimer of this protein. Using CD spectroscopy, the porins investigated were shown to belong to the β-structured proteins. Data of the primary structure and intrinsic protein fluorescence revealed essential differences in localization and microenvironment of tryptophan residues in the porins investigated. Participation of external loops L2 and L6 in the formation of the antigenic structure of OmpC porin from Y. pseudotuberculosis was demonstrated. On the basis of crystal structure of osmoporin from Klebsiella pneumoniae, three-dimensional models of the monomer and trimer of the Y. pseudotuberculosis porin were obtained. Using Web server AGGRESCAN, the localization of protein structure sites with the increased aggregation capability (hot spots) has been deter-mined. It turned out that some of these zones localize in the region of intramonomeric contacts in the porin trimer; however, a large part of them is located on the external surface of the β-barrel. The process of thermal denaturation has been studied and the melting points of the porins were determined. It was found that significant changes in the microenvironment of the indole fluorophores (especially tryptophan residues of spectral class I) took place in the process of the thermodenaturation of the proteins. These changes preceded the irreversible conformational transition observed for the E. coli porin at 77°C and for the Y. pseudotuberculosis porin at 70°C.


Yersinia pseudotuberculosis OmpC porin spatial structure 


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Copyright information

© Pleiades Publishing, Ltd. 2011

Authors and Affiliations

  • O. D. Novikova
    • 1
  • V. A. Khomenko
    • 1
  • V. I. Emelyanenko
    • 2
  • G. N. Likhatskaya
    • 1
  • E. A. Zelepuga
    • 1
  • N. Yu. Kim
    • 1
  • M. P. Isaeva
    • 1
  • O. Yu. Portnyagina
    • 1
  • O. P. Vostrikova
    • 1
  • O. V. Sidorova
    • 1
  • T. F. Solov’eva
    • 1
  1. 1.Pacific Institute of Bioorganic Chemistry, Far East DivisionRussian Academy of SciencesVladivostokRussia
  2. 2.Institute of Theoretical and Experimental BiophysicsRussian Academy of SciencesPushchino, Moscow regionRussia

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