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

SASDSC5 – dark-state long-linker LOV-activated diguanylate cyclase

Sensor domain-containing diguanylate cyclase
MWexperimental 66 kDa
MWexpected 72 kDa
VPorod 115 nm3
log I(s) 6.10×101 6.10×100 6.10×10-1 6.10×10-2
Sensor domain-containing diguanylate cyclase small angle scattering data  s, nm-1
ln I(s)
Sensor domain-containing diguanylate cyclase Guinier plot ln 6.11×101 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
Sensor domain-containing diguanylate cyclase Kratky plot 1.104 0 √3 sRg
p(r)
Sensor domain-containing diguanylate cyclase pair distance distribution function Rg: 2.9 nm 0 Dmax: 11.1 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Sensor domain-containing diguanylate cyclase GASBOR model
Synchrotron SAXS data from solutions of dark-state long-linker LOV-activated diguanylate cyclase in 10 mM Tris, 50 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 0.80 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)

Sensor domain-containing diguanylate cyclase (LadC)
Mol. type   Protein
Organism   Methylotenera sp.
Olig. state   Dimer
Mon. MW   36.2 kDa
 
UniProt   A0A2S5LZS0 (3-314)
Sequence   FASTA