Characterization of murine prion protein

Stefano Da Vela.

SASDHV9 – Murine major prion protein PrPc 23-230

Major prion protein
MWexperimental 21 kDa
MWexpected 23 kDa
VPorod 43 nm3
log I(s) 9.90×10-3 9.90×10-4 9.90×10-5 9.90×10-6
Major prion protein small angle scattering data  s, nm-1
ln I(s)
Major prion protein Guinier plot ln 9.90×10-3 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Major prion protein Kratky plot 1.104 0 3 sRg
p(r)
Major prion protein pair distance distribution function Rg: 2.9 nm 0 Dmax: 9.9 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Major prion protein CORAL model
Major prion protein OTHER [STATIC IMAGE] model

Synchrotron SAXS data from solutions of the major prion protein PrPc (amino acids 23-230) in 10mM HEPES, 150 mM NaCl, pH 7.5 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). Solute concentrations in the nominal range 0.50 - 2.00 mg/ml were 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.

The well-known solubility problems of the full-length PrPc were circumvented by using a buffer containing 150 mM NaCl, in which the protein is partially soluble. The insoluble fraction is pelleted by centrifugation, resulting in a fresh supernatant dominated by the monomer (showing less aggregation than any other condition tested). The SAXS curve for a solution with apparent monomer concentration in the supernatant 1.56 mg/mL was used for the subsequent processing. The structures (amino acids 26-228) were modelled employing 1xyx.pdb for the rigid C-terminal part (amino acids 121-230) and 97 residues as a random loop for the disordered N-terminal part. Shown are the best fit individual model derived from 50 CORAL reconstructions and the spatially-aligned CORAL model cohort (relative to the structured C-terminus) illustrating the spatial-extent/sampling of the disordered N-terminal region of the protein. The individual models and the corresponding fits, as well as a multi-state object with the ensemble as a single pdb file, are made available in the in the full entry zip archive.

Tags: idp
Major prion protein (PrPc)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   22.9 kDa
 
UniProt   P04925 (23-230)
Sequence   FASTA
 
PDB ID   1XYX