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Crystal structure of the fluorescent protein from Dendronephthya sp. in both green and photoconverted red forms.
Acta Crystallogr D Struct Biol 2016; 72(Pt 8):922-32AC

Abstract

The fluorescent protein from Dendronephthya sp. (DendFP) is a member of the Kaede-like group of photoconvertible fluorescent proteins with a His62-Tyr63-Gly64 chromophore-forming sequence. Upon irradiation with UV and blue light, the fluorescence of DendFP irreversibly changes from green (506 nm) to red (578 nm). The photoconversion is accompanied by cleavage of the peptide backbone at the C(α)-N bond of His62 and the formation of a terminal carboxamide group at the preceding Leu61. The resulting double C(α)=C(β) bond in His62 extends the conjugation of the chromophore π system to include imidazole, providing the red fluorescence. Here, the three-dimensional structures of native green and photoconverted red forms of DendFP determined at 1.81 and 2.14 Å resolution, respectively, are reported. This is the first structure of photoconverted red DendFP to be reported to date. The structure-based mutagenesis of DendFP revealed an important role of positions 142 and 193: replacement of the original Ser142 and His193 caused a moderate red shift in the fluorescence and a considerable increase in the photoconversion rate. It was demonstrated that hydrogen bonding of the chromophore to the Gln116 and Ser105 cluster is crucial for variation of the photoconversion rate. The single replacement Gln116Asn disrupts the hydrogen bonding of Gln116 to the chromophore, resulting in a 30-fold decrease in the photoconversion rate, which was partially restored by a further Ser105Asn replacement.

Authors+Show Affiliations

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne, IL 60439, USA.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.Arbuzov Institute of Organic and Physical Chemistry, Russian Academy of Sciences, Kazan, Russian Federation.Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne, IL 60439, USA.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Intramural
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

27487823

Citation

Pletneva, Nadya V., et al. "Crystal Structure of the Fluorescent Protein From Dendronephthya Sp. in Both Green and Photoconverted Red Forms." Acta Crystallographica. Section D, Structural Biology, vol. 72, no. Pt 8, 2016, pp. 922-32.
Pletneva NV, Pletnev S, Pakhomov AA, et al. Crystal structure of the fluorescent protein from Dendronephthya sp. in both green and photoconverted red forms. Acta Crystallogr D Struct Biol. 2016;72(Pt 8):922-32.
Pletneva, N. V., Pletnev, S., Pakhomov, A. A., Chertkova, R. V., Martynov, V. I., Muslinkina, L., ... Pletnev, V. Z. (2016). Crystal structure of the fluorescent protein from Dendronephthya sp. in both green and photoconverted red forms. Acta Crystallographica. Section D, Structural Biology, 72(Pt 8), pp. 922-32. doi:10.1107/S205979831601038X.
Pletneva NV, et al. Crystal Structure of the Fluorescent Protein From Dendronephthya Sp. in Both Green and Photoconverted Red Forms. Acta Crystallogr D Struct Biol. 2016;72(Pt 8):922-32. PubMed PMID: 27487823.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Crystal structure of the fluorescent protein from Dendronephthya sp. in both green and photoconverted red forms. AU - Pletneva,Nadya V, AU - Pletnev,Sergei, AU - Pakhomov,Alexey A, AU - Chertkova,Rita V, AU - Martynov,Vladimir I, AU - Muslinkina,Liya, AU - Dauter,Zbigniew, AU - Pletnev,Vladimir Z, Y1 - 2016/07/13/ PY - 2016/04/07/received PY - 2016/06/27/accepted PY - 2016/8/5/entrez PY - 2016/8/5/pubmed PY - 2017/10/25/medline KW - DendFP KW - DendGFP KW - DendRFP KW - Dendronephthya KW - chromophore KW - fluorescent photoconvertible proteins KW - green fluorescent protein KW - red fluorescent protein KW - structure–function relationships KW - three-dimensional structure KW - β-barrel SP - 922 EP - 32 JF - Acta crystallographica. Section D, Structural biology JO - Acta Crystallogr D Struct Biol VL - 72 IS - Pt 8 N2 - The fluorescent protein from Dendronephthya sp. (DendFP) is a member of the Kaede-like group of photoconvertible fluorescent proteins with a His62-Tyr63-Gly64 chromophore-forming sequence. Upon irradiation with UV and blue light, the fluorescence of DendFP irreversibly changes from green (506 nm) to red (578 nm). The photoconversion is accompanied by cleavage of the peptide backbone at the C(α)-N bond of His62 and the formation of a terminal carboxamide group at the preceding Leu61. The resulting double C(α)=C(β) bond in His62 extends the conjugation of the chromophore π system to include imidazole, providing the red fluorescence. Here, the three-dimensional structures of native green and photoconverted red forms of DendFP determined at 1.81 and 2.14 Å resolution, respectively, are reported. This is the first structure of photoconverted red DendFP to be reported to date. The structure-based mutagenesis of DendFP revealed an important role of positions 142 and 193: replacement of the original Ser142 and His193 caused a moderate red shift in the fluorescence and a considerable increase in the photoconversion rate. It was demonstrated that hydrogen bonding of the chromophore to the Gln116 and Ser105 cluster is crucial for variation of the photoconversion rate. The single replacement Gln116Asn disrupts the hydrogen bonding of Gln116 to the chromophore, resulting in a 30-fold decrease in the photoconversion rate, which was partially restored by a further Ser105Asn replacement. SN - 2059-7983 UR - https://www.unboundmedicine.com/medline/citation/27487823/Crystal_structure_of_the_fluorescent_protein_from_Dendronephthya_sp__in_both_green_and_photoconverted_red_forms_ L2 - http://scripts.iucr.org/cgi-bin/paper?S205979831601038X DB - PRIME DP - Unbound Medicine ER -