Prion Protein-Antibody Complexes Characterized by Chromatography-Coupled Small-Angle X-Ray Scattering.

Carter L, Kim SJ, Schneidman-Duhovny D, Stöhr J, Poncet-Montange G, Weiss TM, Tsuruta H, Prusiner SB, Sali A, Biophys J 109(4):793-805 (2015) Europe PMC

SASDCJ2 – Solution structure of recombinant prion protein (89–230) in complex with Fab-P

Major prion protein
P-Clone Fab, Chimera
MWexperimental 74 kDa
MWexpected 70 kDa
VPorod 106 nm3
log I(s) 1.08×103 1.08×102 1.08×101 1.08×100
Major prion protein P-Clone Fab, Chimera small angle scattering data  s, nm-1
ln I(s)
Major prion protein P-Clone Fab, Chimera Guinier plot ln 1.09×103 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Major prion protein P-Clone Fab, Chimera Kratky plot 1.104 0 √3 sRg
p(r)
Major prion protein P-Clone Fab, Chimera pair distance distribution function Rg: 3.8 nm 0 Dmax: 14.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Major prion protein P-Clone Fab, Chimera PDB (PROTEIN DATA BANK) model
Major prion protein P-Clone Fab, Chimera PDB (PROTEIN DATA BANK) model
log I(s)
 s, nm-1
Major prion protein P-Clone Fab, Chimera PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of recombinant prion protein (89–230) in complex with Fab-P in 20 mM sodium acetate, 150 mM NaCl, pH 5.1 were collected on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSRL, Stanford, CA, USA) using a Rayonix MX225-HE detector at a sample-detector distance of 1.7 m and at a wavelength of λ = 0.113 nm (I(s) vs s, where s = 4πsin θ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1 and 3.7 mg/ml were measured at 9.8°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 and the difference curves were scaled for protein concentration. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Major prion protein (prion)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   22.9 kDa
 
UniProt   P04925 (23-230)
Sequence   FASTA
 
P-Clone Fab, Chimera (Fab-P)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   47.4 kDa
Sequence   FASTA
 
PDB ID   2HH0