Illuminating the inner workings of a natural protein switch: Blue-light sensing in LOV-activated diguanylate cyclases

Vide U, Kasapović D, Fuchs M, Heimböck M, Totaro M, Zenzmaier E, Winkler A, Science Advances 9(31) (2023) DOI

SASDSA5 – dark-state short-linker LOV-activated diguanylate cyclase

Diguanylate cyclase
MWexperimental 72 kDa
MWexpected 67 kDa
VPorod 102 nm3
log I(s) 4.78×101 4.78×100 4.78×10-1 4.78×10-2
Diguanylate cyclase small angle scattering data  s, nm-1
ln I(s)
Diguanylate cyclase Guinier plot ln 4.79×101 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
Diguanylate cyclase Kratky plot 1.104 0 √3 sRg
p(r)
Diguanylate cyclase pair distance distribution function Rg: 3.5 nm 0 Dmax: 13.6 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of dark-state short-linker LOV-activated diguanylate cyclase in 10 mM Tris, 500 mM NaCl, 2 mM MgCl2, pH 8 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus3 2M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.90 mg/ml was measured at 20°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.

Under non-actinic condition (orange light)

Diguanylate cyclase
Mol. type   Protein
Organism   Aquella oligotrophica
Olig. state   Dimer
Mon. MW   33.4 kDa
 
UniProt   A0A2I7N2Y9 (13-304)
Sequence   FASTA