Deciphering Copper Coordination in the Mammalian Prion Protein Amyloidogenic Domain

Salzano G, Brennich M, Mancini G, Tran T, Legname G, D’Angelo P, Giachin G, Biophysical Journal (2020) DOI

SASDEZ7 – C-terminal truncated ovine prion protein (94-234) with ARR polymorphism + Cu(II)

Major prion protein
MWI(0) 15 kDa
MWexpected 16 kDa
VPorod 31 nm3
log I(s) 1.02×101 1.02×100 1.02×10-1 1.02×10-2
Major prion protein small angle scattering data  s, nm-1
ln I(s)
Major prion protein Guinier plot ln 1.02×101 Rg: 2.2 nm 0 (2.2 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: 2.4 nm 0 Dmax: 8.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
C-terminal truncated ovine prion protein (94-234) with ARR polymorphism + Cu(II) Rg histogram Rg, nm
Major prion protein EOM/RANCH model
Synchrotron SAXS data from solutions of the C-terminal truncated ovine prion protein (94-234) with ARR polymorphism in 25 mM ammonium acetate, 250 mM NaCl, 0.001 mM CuSO4, pH 5.5 were collected using size exclusion chromatography SAXS (SEC-SAXS) on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 20 °C using the following parameters: Column type, Superdex 200 5/150 GL (GE Healthcare); Flow rate: 0.15 mL/min; Total acquisition time: 25 min; Injection concentration: 7 mg/mL; Injection volume: 50 μL. The data obtained through the sample elution peak (873-925 1 s frames) were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted and the individual subtracted data sets were scaled and averaged to generate the scattering profile displayed in this entry.

SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

Major prion protein
Mol. type   Protein
Organism   Ovis aries
Olig. state   Monomer
Mon. MW   16.1 kDa
 
UniProt   P23907 (94-234)
Sequence   FASTA