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