Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.

Phototoxic fluorescent proteins represent a sparse group of genetically encoded photosensitizers that could be used for precise light-induced inactivation of target proteins, DNA damage, and cell killing. Only two such GFP-based fluorescent proteins (FPs), KillerRed and its monomeric variant SuperNo...

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Main Authors: Nadya V Pletneva, Vladimir Z Pletnev, Karen S Sarkisyan, Dmitry A Gorbachev, Evgeny S Egorov, Alexander S Mishin, Konstantin A Lukyanov, Zbigniew Dauter, Sergei Pletnev
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2015-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0145740
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author Nadya V Pletneva
Vladimir Z Pletnev
Karen S Sarkisyan
Dmitry A Gorbachev
Evgeny S Egorov
Alexander S Mishin
Konstantin A Lukyanov
Zbigniew Dauter
Sergei Pletnev
author_facet Nadya V Pletneva
Vladimir Z Pletnev
Karen S Sarkisyan
Dmitry A Gorbachev
Evgeny S Egorov
Alexander S Mishin
Konstantin A Lukyanov
Zbigniew Dauter
Sergei Pletnev
author_sort Nadya V Pletneva
collection DOAJ
description Phototoxic fluorescent proteins represent a sparse group of genetically encoded photosensitizers that could be used for precise light-induced inactivation of target proteins, DNA damage, and cell killing. Only two such GFP-based fluorescent proteins (FPs), KillerRed and its monomeric variant SuperNova, were described up to date. Here, we present a crystallographic study of their two orange successors, dimeric KillerOrange and monomeric mKillerOrange, at 1.81 and 1.57 Å resolution, respectively. They are the first orange-emitting protein photosensitizers with a tryptophan-based chromophore (Gln65-Trp66-Gly67). Same as their red progenitors, both orange photosensitizers have a water-filled channel connecting the chromophore to the β-barrel exterior and enabling transport of ROS. In both proteins, Trp66 of the chromophore adopts an unusual trans-cis conformation stabilized by H-bond with the nearby Gln159. This trans-cis conformation along with the water channel was shown to be a key structural feature providing bright orange emission and phototoxicity of both examined orange photosensitizers.
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spelling doaj-art-e36b7701a2794df4983a7c5bab7f0b652025-08-20T02:22:40ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011012e014574010.1371/journal.pone.0145740Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.Nadya V PletnevaVladimir Z PletnevKaren S SarkisyanDmitry A GorbachevEvgeny S EgorovAlexander S MishinKonstantin A LukyanovZbigniew DauterSergei PletnevPhototoxic fluorescent proteins represent a sparse group of genetically encoded photosensitizers that could be used for precise light-induced inactivation of target proteins, DNA damage, and cell killing. Only two such GFP-based fluorescent proteins (FPs), KillerRed and its monomeric variant SuperNova, were described up to date. Here, we present a crystallographic study of their two orange successors, dimeric KillerOrange and monomeric mKillerOrange, at 1.81 and 1.57 Å resolution, respectively. They are the first orange-emitting protein photosensitizers with a tryptophan-based chromophore (Gln65-Trp66-Gly67). Same as their red progenitors, both orange photosensitizers have a water-filled channel connecting the chromophore to the β-barrel exterior and enabling transport of ROS. In both proteins, Trp66 of the chromophore adopts an unusual trans-cis conformation stabilized by H-bond with the nearby Gln159. This trans-cis conformation along with the water channel was shown to be a key structural feature providing bright orange emission and phototoxicity of both examined orange photosensitizers.https://doi.org/10.1371/journal.pone.0145740
spellingShingle Nadya V Pletneva
Vladimir Z Pletnev
Karen S Sarkisyan
Dmitry A Gorbachev
Evgeny S Egorov
Alexander S Mishin
Konstantin A Lukyanov
Zbigniew Dauter
Sergei Pletnev
Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
PLoS ONE
title Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
title_full Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
title_fullStr Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
title_full_unstemmed Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
title_short Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.
title_sort crystal structure of phototoxic orange fluorescent proteins with a tryptophan based chromophore
url https://doi.org/10.1371/journal.pone.0145740
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