Ligands binding to the cellular prion protein

Stefano Da Vela.

SASDLU2 – 2:1 complex between anti prion protein monoclonal IgG 6D11 and recombinant murine prion protein

Major prion protein
Anti-prion protein monoclonal IgG2a 6D11
MWI(0) 206 kDa
MWexpected 168 kDa
VPorod 710 nm3
log I(s) 1.55×10-1 1.55×10-2 1.55×10-3 1.55×10-4
Major prion protein Anti-prion protein monoclonal IgG2a 6D11 small angle scattering data  s, nm-1
ln I(s)
Major prion protein Anti-prion protein monoclonal IgG2a 6D11 Guinier plot ln 1.56×10-1 Rg: 8.1 nm 0 (8.1 nm)-2 s2
(sRg)2I(s)/I(0)
Major prion protein Anti-prion protein monoclonal IgG2a 6D11 Kratky plot 1.104 0 3 sRg
p(r)
Major prion protein Anti-prion protein monoclonal IgG2a 6D11 pair distance distribution function Rg: 7.6 nm 0 Dmax: 24.8 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Major prion protein Anti-prion protein monoclonal IgG2a 6D11 OTHER model

Synchrotron SAXS data from solutions of the 2:1 complex between recombinant murine prion protein and anti prion protein monoclonal IgG 6D11 in PBS, pH 7.4 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 1.80 mg/ml was measured at 20°C. 60 successive 0.095 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.

2:1 molar ratio recombinant murine PrP and 6D11 monoclonal IgG were mixed and incubated 1h on ice. A concentration series in the range 0.3 - 1.8 mg/mL did not show concentration-dependent effects and a representative model of the complex was modeled in SASpy on the 1.8 mg/mL curve taking into account the known epitope and using extended PrP conformations.

Major prion protein (PrPc)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   22.9 kDa
 
UniProt   P04925 (23-230)
Sequence   FASTA
 
Anti-prion protein monoclonal IgG2a 6D11 (6D11)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   145.2 kDa
Sequence   FASTA