Structural basis of GM-CSF and IL-2 sequestration by the viral decoy receptor GIF.

Felix J, Kandiah E, De Munck S, Bloch Y, van Zundert GC, Pauwels K, Dansercoer A, Novanska K, Read RJ, Bonvin AM, Vergauwen B, Verstraete K, Gutsche I, Savvides SN, Nat Commun 7:13228 (2016) Europe PMC

SASDA89 – Complex between ovine GM-CSF and GM-CSF/IL-2 inhibition factor

GM-CSF/IL-2 inhibition factor
Granulocyte-macrophage colony-stimulating factor
MWI(0) 153 kDa
MWexpected 149 kDa
VPorod 231 nm3
log I(s) 1.51×10-1 1.51×10-2 1.51×10-3 1.51×10-4
GM-CSF/IL-2 inhibition factor Granulocyte-macrophage colony-stimulating factor small angle scattering data  s, nm-1
ln I(s)
GM-CSF/IL-2 inhibition factor Granulocyte-macrophage colony-stimulating factor Guinier plot ln 1.51×10-1 Rg: 3.8 nm 0 (3.8 nm)-2 s2
(sRg)2I(s)/I(0)
GM-CSF/IL-2 inhibition factor Granulocyte-macrophage colony-stimulating factor Kratky plot 1.104 0 3 sRg
p(r)
GM-CSF/IL-2 inhibition factor Granulocyte-macrophage colony-stimulating factor pair distance distribution function Rg: 3.8 nm 0 Dmax: 12.4 nm

Data validation


Fits and models


log I(s)
 s, nm-1
GM-CSF/IL-2 inhibition factor Granulocyte-macrophage colony-stimulating factor NONE model

X-ray synchrotron radiation scattering data from solutions of the complex between GM-CSF/IL-2 inhibition factor and ovine Granulocyte-macrophage colony-stimulating factor in 20 mM HEPES 150 mM NaCl were collected on the SWING camera on the storage ring SOLEIL (Saint-Aubin, France) using a CCD AVIEX detector (I(s) vs s, where s = 4π sin θ/λ, where 2θ is the scattering angle). One solute concentration of 10 mg/ml was measured. 10 successive 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 and the different curves were scaled for protein concentration. The purity of the sample was obtained via SEC-SAXS.

Modeling of SAXS data: The crystal structure of the GIF:oGM-CSF complex was used as an input for the Allosmod-FoXS web server to model missing loops, N- and C-termini and N-linked glycosylation. During each Allosmod-FoXS run, data up to a scattering angle of 0.5 Å-1 was used. Fits to the experimental SAXS data were calculated using the FoXS software.

GM-CSF/IL-2 inhibition factor
Mol. type   Protein
Organism   Orf virus
Olig. state   Tetramer
Mon. MW   29.9 kDa
 
UniProt   Q9J5U5
Sequence   FASTA
 
Granulocyte-macrophage colony-stimulating factor
Mol. type   Protein
Organism   Ovis aries
Olig. state   Monomers
Mon. MW   14.4 kDa
 
UniProt   P28773 (18-144)
Sequence   FASTA