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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. https://proteinensemble.org/entries/PED00219
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s, nm-1
![Major prion protein OTHER [STATIC IMAGE] model](/media//pdb_file/SASDHV9_fit1_model2.png "Static model image")
