1. Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein
- Author
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Röllen, Katrin, Granzin, Joachim, Batra-Safferling, Renu, and Stadler, Andreas Maximilian
- Subjects
Photoreceptors ,Sensory Receptors ,Flavin Mononucleotide ,Social Sciences ,lcsh:Medicine ,Crystallography, X-Ray ,Photoreceptors, Microbial ,Biochemistry ,Protein Structure, Secondary ,Small-Angle Scattering ,Scattering ,Spectrum Analysis Techniques ,X-Ray Diffraction ,Animal Cells ,Relaxation Time ,Psychology ,Post-Translational Modification ,lcsh:Science ,Neurons ,Crystallography ,Flavin ,Physics ,Absorption Spectroscopy ,Condensed Matter Physics ,Chemistry ,Physical Sciences ,Crystal Structure ,Sensory Perception ,ddc:500 ,Cellular Types ,Research Article ,Signal Transduction ,Chemical physics ,Research and Analysis Methods ,Bacterial Proteins ,Protein Domains ,Scattering, Small Angle ,Solid State Physics ,ddc:610 ,Relaxation (Physics) ,Flavoproteins ,Pseudomonas putida ,fungi ,lcsh:R ,Biology and Life Sciences ,Proteins ,Afferent Neurons ,Dimers (Chemical physics) ,Cell Biology ,Oxygen ,Kinetics ,Cellular Neuroscience ,Ultraviolet-Visible Spectroscopy ,Spectrophotometry, Ultraviolet ,lcsh:Q ,Neuroscience - Abstract
PLOS ONE 13(7), e0200746 - (2018). doi:10.1371/journal.pone.0200746, 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 τREC 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., Published by PLOS, San Francisco, California, US
- Published
- 2018