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Three-dimensional structure of a pH-dependent fluorescent protein WasCFP with a tryptophan based deprotonated chromophore

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Abstract

WasCFP, a pH-dependent green fluorescent protein with a tryptophan-based chromophore (Thr65-Trp66-Gly67) in anionic state, was designed from a cyan precursor mCerulean. In this study, the three-dimensional structure of WasCFP has been determined by an X-ray method at pH 5.5, pH 8.0 and pH 10.0, with a resolution of 1.14, 1.25 and 1.5 Å, respectively. We show that changes in the acidity of the media are accompanied by a synchronous change of the side chain conformations of the residues in the near-chromophore environment. Subsequent changes in the local H-bond network interacting with the chromophore lead to considerable alterations in the protein spectral properties as a consequence of reversible processes of ionization-protonation of the Trp chromophore. These experimental results have been supported by quantum chemistry calculations.

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Abbreviations

GFP:

green fluorescent protein

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Russia

    V. Z. Pletnev, N. V. Pletneva, R. G. Efremov, E. A. Goryacheva, I. V. Artemyev, S. F. Arkhipova, K. S. Sarkisyan, A. S. Mishin & K. A. Lukyanov

  2. Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne, IL, 60439, USA

    S. V. Pletnev

Authors
  1. V. Z. Pletnev
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  2. N. V. Pletneva
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  3. R. G. Efremov
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  4. E. A. Goryacheva
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  5. I. V. Artemyev
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  6. S. F. Arkhipova
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  7. K. S. Sarkisyan
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  8. A. S. Mishin
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  9. K. A. Lukyanov
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Correspondence to V. Z. Pletnev.

Additional information

Original Russian Text © V.Z. Pletnev, N.V. Pletneva, R.G. Efremov, E.A. Goryacheva, I.V. Artemyev, S.F. Arkhipova, K.S. Sarkisyan, A.S. Mishin, K.A. Lukyanov, S.V. Pletnev, 2016, published in Bioorganicheskaya Khimiya, 2016, Vol. 42, No. 6, pp. 675–682.

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Pletnev, V.Z., Pletneva, N.V., Efremov, R.G. et al. Three-dimensional structure of a pH-dependent fluorescent protein WasCFP with a tryptophan based deprotonated chromophore. Russ J Bioorg Chem 42, 612–618 (2016). https://doi.org/10.1134/S1068162016050149

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  • DOI: https://doi.org/10.1134/S1068162016050149

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