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Isolation and characterization of recombinant OmpF-like porin from the Yersinia pseudotuberculosis outer membrane

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Abstract

The encoding sequence of the pore-forming OmpF-like protein from the Yersinia pseudotuberculosis outer membrane was cloned and expressed in Escherichia coli cells. Conditions for isolation and refolding of recombinant monomer and porin trimer were selected. Their spatial structures were characterized by the intrinsic protein fluorescence and CD spectroscopy. It was shown that recombinant porins are similar in the composition of secondary structure elements to isolated porins, but have a considerably less compact tertiary structure. The pore-forming activities of the recombinant proteins are similar to those of Y. pseudotuberculosis native porins.

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Abbreviations

BLM:

bilayer lipid membrane

TLT and TSM:

thermolabile trimer and thermostable monomer forms of Y. pseudotuberculosis porine

DOC:

sodium deoxycholate

IB:

inclusion bodies

IPTG:

isopropyl β-D-thiogalactopyranoside

LPS:

lipopolysaccharide

OM:

outer membrane

PMSF:

phenylmethylsulfonyl fluoride

RP:

recombinant protein

Zw.3–14:

N-tetradecyl-N,N-dimethyl-3-ammonium 1-propanesulfonate

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Authors and Affiliations

  1. Pacific Institute of Bioorganic Chemistry, Far East Division, Russian Academy of Sciences, pr. Stoletiya Vladivostoka 159, Vladivostok, 690022, Russia

    V. A. Khomenko, O. Yu. Portnyagina, O. D. Novikova, M. P. Isaeva, N. Yu. Kim, G. N. Likhatskaya, O. P. Vostrikova & T. F. Solov’eva

Authors
  1. V. A. Khomenko
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  2. O. Yu. Portnyagina
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  3. O. D. Novikova
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  4. M. P. Isaeva
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  5. N. Yu. Kim
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  6. G. N. Likhatskaya
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  7. O. P. Vostrikova
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  8. T. F. Solov’eva
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Corresponding author

Correspondence to O. Yu. Portnyagina.

Additional information

Original Russian Text © V.A. Khomenko, O.Yu. Portnyagina, O.D. Novikova, M.P. Isaeva, N.Yu. Kim, G.N. Likhatskaya, O.P. Vostrikova, T.F. Solov’eva, 2008, published in Bioorganicheskaya Khimiya, 2008, Vol. 34, No. 2, pp. 177–184.

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Khomenko, V.A., Portnyagina, O.Y., Novikova, O.D. et al. Isolation and characterization of recombinant OmpF-like porin from the Yersinia pseudotuberculosis outer membrane. Russ J Bioorg Chem 34, 162–168 (2008). https://doi.org/10.1134/S1068162008020040

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  • DOI: https://doi.org/10.1134/S1068162008020040

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