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Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2
A structural analysis of the recently developed orange fluorescent proteins with novel phenotypes, LSSmOrange (λ(ex)/λ(em) at 437/572 nm), PSmOrange (λ(ex)/λ(em) at 548/565 nm and for photoconverted form at 636/662 nm) and PSmOrange2 (λ(ex)/λ(em) at 546/561 nm and for photoconverted form at 619/651...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068994/ https://www.ncbi.nlm.nih.gov/pubmed/24960050 http://dx.doi.org/10.1371/journal.pone.0099136 |
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author | Pletnev, Sergei Shcherbakova, Daria M. Subach, Oksana M. Pletneva, Nadya V. Malashkevich, Vladimir N. Almo, Steven C. Dauter, Zbigniew Verkhusha, Vladislav V. |
author_facet | Pletnev, Sergei Shcherbakova, Daria M. Subach, Oksana M. Pletneva, Nadya V. Malashkevich, Vladimir N. Almo, Steven C. Dauter, Zbigniew Verkhusha, Vladislav V. |
author_sort | Pletnev, Sergei |
collection | PubMed |
description | A structural analysis of the recently developed orange fluorescent proteins with novel phenotypes, LSSmOrange (λ(ex)/λ(em) at 437/572 nm), PSmOrange (λ(ex)/λ(em) at 548/565 nm and for photoconverted form at 636/662 nm) and PSmOrange2 (λ(ex)/λ(em) at 546/561 nm and for photoconverted form at 619/651 nm), is presented. The obtained crystallographic structures provide an understanding of how the ensemble of a few key mutations enabled special properties of the orange FPs. While only a single Ile161Asp mutation, enabling excited state proton transfer, is critical for LSSmOrange, other substitutions provide refinement of its special properties and an exceptional 120 nm large Stokes shift. Similarly, a single Gln64Leu mutation was sufficient to cause structural changes resulting in photoswitchability of PSmOrange, and only one additional substitution (Phe65Ile), yielding PSmOrange2, was enough to greatly decrease the energy of photoconversion and increase its efficiency of photoswitching. Fluorescence of photoconverted PSmOrange and PSmOrange2 demonstrated an unexpected bathochromic shift relative to the fluorescence of classic red FPs, such as DsRed, eqFP578 and zFP574. The structural changes associated with this fluorescence shift are of considerable value for the design of advanced far-red FPs. For this reason the chromophore transformations accompanying photoconversion of the orange FPs are discussed. |
format | Online Article Text |
id | pubmed-4068994 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-40689942014-06-27 Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 Pletnev, Sergei Shcherbakova, Daria M. Subach, Oksana M. Pletneva, Nadya V. Malashkevich, Vladimir N. Almo, Steven C. Dauter, Zbigniew Verkhusha, Vladislav V. PLoS One Research Article A structural analysis of the recently developed orange fluorescent proteins with novel phenotypes, LSSmOrange (λ(ex)/λ(em) at 437/572 nm), PSmOrange (λ(ex)/λ(em) at 548/565 nm and for photoconverted form at 636/662 nm) and PSmOrange2 (λ(ex)/λ(em) at 546/561 nm and for photoconverted form at 619/651 nm), is presented. The obtained crystallographic structures provide an understanding of how the ensemble of a few key mutations enabled special properties of the orange FPs. While only a single Ile161Asp mutation, enabling excited state proton transfer, is critical for LSSmOrange, other substitutions provide refinement of its special properties and an exceptional 120 nm large Stokes shift. Similarly, a single Gln64Leu mutation was sufficient to cause structural changes resulting in photoswitchability of PSmOrange, and only one additional substitution (Phe65Ile), yielding PSmOrange2, was enough to greatly decrease the energy of photoconversion and increase its efficiency of photoswitching. Fluorescence of photoconverted PSmOrange and PSmOrange2 demonstrated an unexpected bathochromic shift relative to the fluorescence of classic red FPs, such as DsRed, eqFP578 and zFP574. The structural changes associated with this fluorescence shift are of considerable value for the design of advanced far-red FPs. For this reason the chromophore transformations accompanying photoconversion of the orange FPs are discussed. Public Library of Science 2014-06-24 /pmc/articles/PMC4068994/ /pubmed/24960050 http://dx.doi.org/10.1371/journal.pone.0099136 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. |
spellingShingle | Research Article Pletnev, Sergei Shcherbakova, Daria M. Subach, Oksana M. Pletneva, Nadya V. Malashkevich, Vladimir N. Almo, Steven C. Dauter, Zbigniew Verkhusha, Vladislav V. Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title | Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title_full | Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title_fullStr | Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title_full_unstemmed | Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title_short | Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2 |
title_sort | orange fluorescent proteins: structural studies of lssmorange, psmorange and psmorange2 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068994/ https://www.ncbi.nlm.nih.gov/pubmed/24960050 http://dx.doi.org/10.1371/journal.pone.0099136 |
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