Argonaute protein SAXS investigation

lirong zheng.

SASDPN2 – apo-Pyrococcus furiosus argonaute protein at 95 °C

Piwi domain-containing protein
MWexperimental 81 kDa
MWexpected 91 kDa
VPorod 187 nm3
log I(s) 1.16×10-1 1.16×10-2 1.16×10-3 1.16×10-4
Piwi domain-containing protein small angle scattering data  s, nm-1
ln I(s)
Piwi domain-containing protein Guinier plot ln 1.17×10-1 Rg: 3.2 nm 0 (3.2 nm)-2 s2
(sRg)2I(s)/I(0)
Piwi domain-containing protein Kratky plot 1.104 0 √3 sRg
p(r)
Piwi domain-containing protein pair distance distribution function Rg: 3.4 nm 0 Dmax: 9.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Piwi domain-containing protein GASBOR model
log I(s)
 s, nm-1
Piwi domain-containing protein PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of apo-Pyrococcus furiosus argonaute protein in 20 mM Tris–HCl, 250 mM NaCl, 2mM DTT, pH 8 were collected on the BL19U2 beam line at the Shanghai Synchrotron Radiation Facility (SSRF, Shanghai, China) using a Pilatus 1M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.30 mg/ml was measured at 95°C. 20 successive 0.500 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.

Substitute errors have been used for the ab initio GASBOR modelling (+/-1.0 on each data point).

Piwi domain-containing protein (apo-pfago)
Mol. type   Protein
Organism   Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Olig. state   Monomer
Mon. MW   91.3 kDa
 
UniProt   Q8U3D2 (5-770)
Sequence   FASTA
 
PDB ID   1Z26