Three-dimensional structure of a pH-dependent fluorescent protein WasCFP with a tryptophan based deprotonated chromophore
- Authors: Pletnev V.Z.1, Pletneva N.V.1, Efremov R.G.1, Goryacheva E.A.1, Artemyev I.V.1, Arkhipova S.F.1, Sarkisyan K.S.1, Mishin A.S.1, Lukyanov K.A.1, Pletnev S.V.2
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Affiliations:
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
- Synchrotron Radiation Research Section
- Issue: Vol 42, No 6 (2016)
- Pages: 612-618
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228182
- DOI: https://doi.org/10.1134/S1068162016050149
- ID: 228182
Cite item
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.
About the authors
V. Z. Pletnev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Author for correspondence.
Email: vzpletnev@gmail.com
Russian Federation, Moscow
N. V. Pletneva
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
R. G. Efremov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
E. A. Goryacheva
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
I. V. Artemyev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
S. F. Arkhipova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
K. S. Sarkisyan
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
A. S. Mishin
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
K. A. Lukyanov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Russian Federation, Moscow
S. V. Pletnev
Synchrotron Radiation Research Section
Email: vzpletnev@gmail.com
United States, Argonne, IL, 60439