Differentiating the solution structures and stability of transthyretin tetramer complexed with tolcapone and tafamidis using SEC-SWAXS and NMR

Orion Shih.

SASDXW3 – Transthyretin tetramer

Transthyretin
MWexperimental 59 kDa
MWexpected 55 kDa
VPorod 92 nm3
log I(s) 7.47×10-2 7.47×10-3 7.47×10-4 7.47×10-5
Transthyretin small angle scattering data  s, nm-1
ln I(s)
Transthyretin Guinier plot ln 7.47×10-2 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Transthyretin Kratky plot 1.104 0 √3 sRg
p(r)
Transthyretin pair distance distribution function Rg: 2.5 nm 0 Dmax: 7.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Transthyretin COOT model
Synchrotron SAXS data from solutions of transthyretin in 20 mM Tris, 50 mM NaCl, pH 7 were collected on the 13A beam line at the Taiwan Photon Source, NSRRC (Hsinchu, Taiwan) using a Eiger X 1M and Eiger X 9M detector at a sample-detector distance of 0.3 m (Eiger X 1M) and 2.5 m (Eiger X 9M) and at a wavelength of f λ = 0.08265 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 26°C. Eight successive 2 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.

Transthyretin (TTR)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   13.8 kDa
 
UniProt   P02766 (21-147)
Sequence   FASTA