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I87E Mutation Prevents Barstar Dimerization

Russian Journal of Bioorganic Chemistry volume 30, pages 577–581 (2004)Cite this article

Abstract

The C40,82A;I87E mutant of barstar, an intracellular inhibitor of the ribonuclease barnase from Bacillus amyloliquefaciens, was obtained, and its physicochemical properties were studied. It was produced as a fusion protein with thioredoxin and then cleaved from this by EKmax enterokinase. The mutant was shown by NMR to retain the spatial structure of the wild-type protein but, in contrast to barstar, does not form the homodimers characteristic of barstar in aqueous solution. The mutant protein binds barnase with the dissociation constant (6.6 ± 1.1) × 10–11 M and exhibits other physicochemical properties similar to those of the wild-type barstar. This allows the use of C40,82A;I87E mutant instead of wild-type barstar in the investigations where the protein dimerization is undesirable.

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    D. S. Korchuganov, A. A. Schulga, Ya. S. Ermolyuk, M. Ya. Reibarkh, S. B. Nolde, A. S. Arseniev & M. P. Kirpichnikov

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow, 119991, Russia

    V. A. Mitkevich & A. A. Makarov

  3. The Center for Medical Studies at Moscow, University of Oslo, ul. Vavilova 34/5, Moscow, 119991, Russia

    V. A. Mitkevich

Authors
  1. D. S. Korchuganov
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  2. A. A. Schulga
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  3. Ya. S. Ermolyuk
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  4. V. A. Mitkevich
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  5. M. Ya. Reibarkh
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  6. S. B. Nolde
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  7. A. A. Makarov
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  8. A. S. Arseniev
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  9. M. P. Kirpichnikov
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Korchuganov, D.S., Schulga, A.A., Ermolyuk, Y.S. et al. I87E Mutation Prevents Barstar Dimerization. Russian Journal of Bioorganic Chemistry 30, 577–581 (2004). https://doi.org/10.1023/B:RUBI.0000049775.11808.a1

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  • DOI: https://doi.org/10.1023/B:RUBI.0000049775.11808.a1

  • barnase
  • barstar
  • protein dimerization