Structural and enzymatic characterisation of the Type III effector NopAA (=GunA) from Sinorhizobium fredii USDA257 reveals a Xyloglucan hydrolase activity.

Dorival J, Philys S, Giuntini E, Brailly R, de Ruyck J, Czjzek M, Biondi E, Bompard C, Sci Rep 10(1):9932 (2020) Europe PMC

SASDHG4 – NopAA, a type three effector from Sinorhizobium fredii USDA257 whith xyloglucanase activity

Type III effector NopAA
MWexperimental 31 kDa
MWexpected 31 kDa
VPorod 38 nm3
log I(s) 2.33×10-2 2.33×10-3 2.33×10-4 2.33×10-5
Type III effector NopAA small angle scattering data  s, nm-1
ln I(s)
Type III effector NopAA Guinier plot ln 2.33×10-2 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Type III effector NopAA Kratky plot 1.104 0 3 sRg
p(r)
Type III effector NopAA pair distance distribution function Rg: 2.6 nm 0 Dmax: 9.9 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Type III effector NopAA ALLOSMOD model

Synchrotron SAXS data from solutions of NopAA in PBS, 150 mM NaCl, 10% glycerol, pH 7.4 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a wavelength of λ = 1.033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 20 mg/ml was injected at a 0.20 ml/min flow rate onto a Agilent Bio SEC-3, 150 Å column at 15°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Type III effector NopAA (NopAA)
Mol. type   Protein
Organism   Sinorhizobium fredii USDA257
Olig. state   Monomer
Mon. MW   30.8 kDa
 
UniProt   M4PUR5 (1-268)
Sequence   FASTA