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Russian Journal of Bioorganic Chemistry

, Volume 42, Issue 6, pp 612–618 | Cite as

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

  • 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
Article
  • 53 Downloads

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.

Keywords

crystal structure green fluorescent protein WasCFP Trp based chromophore Trp anionic form 

Abbreviations

GFP

green fluorescent protein

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Copyright information

© Pleiades Publishing, Ltd. 2016

Authors and Affiliations

  • V. Z. Pletnev
    • 1
  • N. V. Pletneva
    • 1
  • R. G. Efremov
    • 1
  • E. A. Goryacheva
    • 1
  • I. V. Artemyev
    • 1
  • S. F. Arkhipova
    • 1
  • K. S. Sarkisyan
    • 1
  • A. S. Mishin
    • 1
  • K. A. Lukyanov
    • 1
  • S. V. Pletnev
    • 2
  1. 1.Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of SciencesMoscowRussia
  2. 2.Synchrotron Radiation Research SectionMacromolecular Crystallography Laboratory, National Cancer InstituteArgonneUSA

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