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Title: Crystal structure of phototoxic orange fluorescent proteins with α 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 SuperNova, were described up to date. We present a crystallographic study of their two orange successors, dimeric KillerOrange and monomeric mKiller-Orange, 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.
Authors:
 [1] ;  [1] ;  [1] ;  [2] ;  [1] ;  [3] ;  [3] ;  [4] ;  [5]
  1. Russian Academy of Sciences (RAS), Moscow (Russian Federation)
  2. Russian Academy of Sciences (RAS), Moscow (Russian Federation); Moscow State Univ., Moscow (Russian Federation)
  3. Russian Academy of Sciences (RAS), Moscow (Russian Federation); Nizhny Novgorod State Medical Academy, Nizhny Novgorod (Russian Federation)
  4. National Cancer Insitute, Argonne, IL (United States)
  5. National Cancer Insitute, Argonne, IL (United States); Leidos Biomedical Research, Inc., Argonne, IL (United States)
Publication Date:
Type:
Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 10; Journal Issue: 12; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1239529

Pletneva, Nadya V., Pletnev, Vladimir Z., Sarkisyan, Karen S., Gorbachev, Dmitry A., Egorov, Evgeny S., Mishin, Alexander S., Lukyanov, Konstantin A., Dauter, Zbigniew, and Pletnev, Sergei. Crystal structure of phototoxic orange fluorescent proteins with α tryptophan-based chromophore. United States: N. p., Web. doi:10.1371/journal.pone.0145740.
Pletneva, Nadya V., Pletnev, Vladimir Z., Sarkisyan, Karen S., Gorbachev, Dmitry A., Egorov, Evgeny S., Mishin, Alexander S., Lukyanov, Konstantin A., Dauter, Zbigniew, & Pletnev, Sergei. Crystal structure of phototoxic orange fluorescent proteins with α tryptophan-based chromophore. United States. doi:10.1371/journal.pone.0145740.
Pletneva, Nadya V., Pletnev, Vladimir Z., Sarkisyan, Karen S., Gorbachev, Dmitry A., Egorov, Evgeny S., Mishin, Alexander S., Lukyanov, Konstantin A., Dauter, Zbigniew, and Pletnev, Sergei. 2015. "Crystal structure of phototoxic orange fluorescent proteins with α tryptophan-based chromophore". United States. doi:10.1371/journal.pone.0145740. https://www.osti.gov/servlets/purl/1239529.
@article{osti_1239529,
title = {Crystal structure of phototoxic orange fluorescent proteins with α tryptophan-based chromophore},
author = {Pletneva, Nadya V. and Pletnev, Vladimir Z. and Sarkisyan, Karen S. and Gorbachev, Dmitry A. and Egorov, Evgeny S. and Mishin, Alexander S. and Lukyanov, Konstantin A. and Dauter, Zbigniew and Pletnev, Sergei},
abstractNote = {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. We present a crystallographic study of their two orange successors, dimeric KillerOrange and monomeric mKiller-Orange, 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.},
doi = {10.1371/journal.pone.0145740},
journal = {PLoS ONE},
number = 12,
volume = 10,
place = {United States},
year = {2015},
month = {12}
}