Experimental Evidence of Intrinsic Disorder and Amyloid Formation by the Henipavirus W Proteins

Pesce G, Gondelaud F, Ptchelkine D, Nilsson J, Bignon C, Cartalas J, Fourquet P, Longhi S, International Journal of Molecular Sciences 23(2):923 (2022) DOI

SASDLK9 – Hendra virus W protein in 1 M urea, 5 mM DTT

Protein W
MWexperimental 53 kDa
MWexpected 53 kDa
VPorod 338 nm3
log I(s) 2.98×10-2 2.98×10-3 2.98×10-4 2.98×10-5
Protein W small angle scattering data  s, nm-1
ln I(s)
Protein W Guinier plot ln 2.98×10-2 Rg: 7.2 nm 0 (7.2 nm)-2 s2
(sRg)2I(s)/I(0)
Protein W Kratky plot 1.104 0 √3 sRg
p(r)
Protein W pair distance distribution function Rg: 7.4 nm 0 Dmax: 24 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Hendra virus W protein in 1 M urea, 5 mM DTT Rg histogram Rg, nm
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Protein W EOM/RANCH model
Synchrotron SAXS data from solutions of Hendra virus W protein in 20 mM HEPES, 150 mM NaCl, 1 M urea, 5 mM DTT, pH 7 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a Eiger 4M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 70.00 μl sample at 2.4 mg/ml was injected at a 0.20 ml/min flow rate onto a Agilent Bio SEC-3, 300 Å column at 20°C. 600 successive 0.990 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.

Molecular weight of the Hendra W protein has been determined by MALDI-TOF to 52.577 kDa. Hendra virus W protein expressed as a recombinant protein possessing a N-terminal 6xHis tag PED: https://proteinensemble.org/PED00204

Tags: idp PED
Protein W (HeVW)
Mol. type   Protein
Organism   Hendra virus (isolate Horse/Autralia/Hendra/1994)
Olig. state   Monomer
Mon. MW   52.6 kDa
 
UniProt   P0C1C6 (2-448)
Sequence   FASTA
 
PED ID   PED00204