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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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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. |
format | Online Article Text |
id | pubmed-6047819 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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