Small angle X-ray scattering as a complementary tool for high-throughput structural studies.

Grant TD, Luft JR, Wolfley JR, Tsuruta H, Martel A, Montelione GT, Snell EH, Biopolymers 2011 Aug;95(8):517-30 PubMed

SASDCF6 – HIT family hydrolase protein from Vibrio fischeri. Northeast Structural Genomics Consortium target id VfR176

HIT family hydrolase
MWexperimental 34.8 kDa
MWexpected 34 kDa
VPorod 57.8 nm3
log I(s) 1.06×103 1.06×102 1.06×101 1.06×100
HIT family hydrolase small angle scattering data  s, nm-1
ln I(s)
HIT family hydrolase Guinier plot ln 1.06×103 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
HIT family hydrolase Kratky plot 1.104 0 3 sRg
p(r)
HIT family hydrolase pair distance distribution function Rg: 2.1 nm 0 Dmax: 7.2 nm

Experimental data validation


Fits and models


log I(s)
 s, nm-1
HIT family hydrolase PDB model
log I(s)
 s, nm-1
HIT family hydrolase DAMFILT model
Synchrotron SAXS data from solutions of HIT family hydrolase protein from Vibrio fischeri. Northeast Structural Genomics Consortium target id VfR176 in 5 mM DTT 100 mM NaCl 10 mM Tris-HCl 0.02 % NaN3, pH 7.5 were collected on the BL4-2 camera on the storage ring Stanford Synchrotron Radiation Lightsource (SSRL) (Stanford, CA, USA). Using a Rayonix MX225-HE detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.13 nm (s = 4π sin θ/λ, where 2θ is the scattering angle). Solute concentrations ranging between 2.1 and 6.3 mg/ml were measured at 20°C. 20 successive 1 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 and the different curves were scaled for protein concentration.

HIT family hydrolase
Mol. type   Protein
Organism   Vibrio fischeri (strain ATCC 700601 / ES114)
Olig. state   Dimer
Mon. MW   17.0 kDa
 
UniProt   Q5E3V1
Sequence   FASTA
 
PDB code   3I24