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TitleSmall-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein.
Journal, issue, pagesPLoS One, Vol. 13, Issue 7, Page e0200746, Year 2018
Publish dateJul 16, 2018
AuthorsKatrin Röllen / Joachim Granzin / Renu Batra-Safferling / Andreas Maximilian Stadler /
PubMed AbstractLight, 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, 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.
External linksPLoS One / PubMed:30011332 / PubMed Central
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution2.04 Å
Structure data

SASDDE6:
Photoreceptor sensory box 1 light-state (SB1-LOV-R66I)
Method: SAXS/SANS

SASDDG6:
Photoreceptor sensory box 1 dark-state (SB1-LOV-R66I)
Method: SAXS/SANS

PDB-6gg9:
Crystal structures of a short blue light photoreceptor protein PpSB1-LOV mutant (dark state) - R61H/R66I
Method: X-RAY DIFFRACTION / Resolution: 2.04 Å

Chemicals

ChemComp-FMN:
FLAVIN MONONUCLEOTIDE

ChemComp-HOH:
WATER

Source
  • Pseudomonas putida (strain atcc 47054 / dsm 6125 / ncimb 11950 / kt2440) (bacteria)
  • pseudomonas putida (bacteria)
KeywordsSIGNALING PROTEIN / blue light photoreceptor / Sensory box protein

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