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Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein

Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue-light induced conformational transition of the dimeric photoreceptor PpSB1-LOV-R66I from Pseudomonas...

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Autores principales: Röllen, Katrin, Granzin, Joachim, Batra-Safferling, Renu, Stadler, Andreas Maximilian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6047819/
https://www.ncbi.nlm.nih.gov/pubmed/30011332
http://dx.doi.org/10.1371/journal.pone.0200746
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author Röllen, Katrin
Granzin, Joachim
Batra-Safferling, Renu
Stadler, Andreas Maximilian
author_facet Röllen, Katrin
Granzin, Joachim
Batra-Safferling, Renu
Stadler, Andreas Maximilian
author_sort Röllen, Katrin
collection PubMed
description Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue-light induced conformational transition of the dimeric photoreceptor PpSB1-LOV-R66I from Pseudomonas putida in solution by using small-angle X-ray scattering (SAXS). SAXS experiments of the fully populated light- and dark-states under steady-state conditions revealed significant structural differences between the two states that are in agreement with the known structures determined by crystallography. We followed the transition from the light- to the dark-state by using SAXS measurements in real-time. A two-state model based on the light- and dark-state conformations could describe the measured time-course SAXS data with a relaxation time τ(REC) of ~ 34 to 35 min being larger than the recovery time found with UV/vis spectroscopy. Unlike the flavin chromophore-based UV/vis method that is sensitive to the local chromophore environment in flavoproteins, SAXS-based assay depends on protein conformational changes and provides with an alternative to measure the recovery kinetics.
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spelling pubmed-60478192018-07-26 Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein Röllen, Katrin Granzin, Joachim Batra-Safferling, Renu Stadler, Andreas Maximilian PLoS One Research Article Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue-light induced conformational transition of the dimeric photoreceptor PpSB1-LOV-R66I from Pseudomonas putida in solution by using small-angle X-ray scattering (SAXS). SAXS experiments of the fully populated light- and dark-states under steady-state conditions revealed significant structural differences between the two states that are in agreement with the known structures determined by crystallography. We followed the transition from the light- to the dark-state by using SAXS measurements in real-time. A two-state model based on the light- and dark-state conformations could describe the measured time-course SAXS data with a relaxation time τ(REC) of ~ 34 to 35 min being larger than the recovery time found with UV/vis spectroscopy. Unlike the flavin chromophore-based UV/vis method that is sensitive to the local chromophore environment in flavoproteins, SAXS-based assay depends on protein conformational changes and provides with an alternative to measure the recovery kinetics. Public Library of Science 2018-07-16 /pmc/articles/PMC6047819/ /pubmed/30011332 http://dx.doi.org/10.1371/journal.pone.0200746 Text en © 2018 Röllen et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Röllen, Katrin
Granzin, Joachim
Batra-Safferling, Renu
Stadler, Andreas Maximilian
Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title_full Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title_fullStr Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title_full_unstemmed Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title_short Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
title_sort small-angle x-ray scattering study of the kinetics of light-dark transition in a lov protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6047819/
https://www.ncbi.nlm.nih.gov/pubmed/30011332
http://dx.doi.org/10.1371/journal.pone.0200746
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