Three-dimensional structure of a pH-dependent fluorescent protein WasCFP with a tryptophan based deprotonated chromophore
- Autores: Pletnev V.1, Pletneva N.1, Efremov R.1, Goryacheva E.1, Artemyev I.1, Arkhipova S.1, Sarkisyan K.1, Mishin A.1, Lukyanov K.1, Pletnev S.2
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Afiliações:
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
- Synchrotron Radiation Research Section
- Edição: Volume 42, Nº 6 (2016)
- Páginas: 612-618
- Seção: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228182
- DOI: https://doi.org/10.1134/S1068162016050149
- ID: 228182
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Resumo
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.
Sobre autores
V. Pletnev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Autor responsável pela correspondência
Email: vzpletnev@gmail.com
Rússia, Moscow
N. Pletneva
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
R. Efremov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
E. Goryacheva
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
I. Artemyev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
S. Arkhipova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
K. Sarkisyan
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
A. Mishin
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
K. Lukyanov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Email: vzpletnev@gmail.com
Rússia, Moscow
S. Pletnev
Synchrotron Radiation Research Section
Email: vzpletnev@gmail.com
Estados Unidos da América, Argonne, IL, 60439