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

Röllen K, Granzin J, Batra-Safferling R, Stadler AM, PLoS One 13(7):e0200746 (2018) Europe PMC

SASDDG6 – Photoreceptor sensory box 1 dark-state (SB1-LOV-R66I)

Sensory box protein dark-state
MWI(0) 42 kDa
MWexpected 37 kDa
VPorod 57 nm3
log I(s) 4.22×101 4.22×100 4.22×10-1 4.22×10-2
Sensory box protein dark-state small angle scattering data  s, nm-1
ln I(s)
Sensory box protein dark-state Guinier plot ln 4.22×101 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Sensory box protein dark-state Kratky plot 1.104 0 √3 sRg
p(r)
Sensory box protein dark-state pair distance distribution function Rg: 2.5 nm 0 Dmax: 8.0 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Photoreceptor sensory box 1 dark-state (SB1-LOV-R66I) Rg histogram Rg, nm
Sensory box protein dark-state EOM/RANCH model
Sensory box protein dark-state EOM/RANCH model
Sensory box protein dark-state EOM/RANCH model
Sensory box protein dark-state EOM/RANCH model
Sensory box protein dark-state EOM/RANCH model
Sensory box protein dark-state EOM/RANCH model
log I(s)
 s, nm-1
Sensory box protein dark-state DAMMIN model
log I(s)
 s, nm-1
Sensory box protein dark-state CORAL model
Synchrotron SAXS data from solutions of the dark-state photoreceptor sensory box 1 protein (SB1-LOV-R66I) in 10mM Tris, 10 mM NaCl, pH 7, were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2.5 and 6.1 mg/ml were measured at 10°C. 10 successive 1 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Sensory box protein dark-state
Mol. type   Protein
Organism   Pseudomonas putida
Olig. state   Dimer
Mon. MW   18.6 kDa
 
UniProt   Q88E39
Sequence   FASTA