Improving data quality and expanding BioSAXS experiments to low-molecular-weight and low-concentration protein samples.

Castellví A, Pascual-Izarra C, Crosas E, Malfois M, Juanhuix J, Acta Crystallogr D Struct Biol 76(Pt 10):971-981 (2020) Europe PMC

SASDJJ3 – SH3 domain of alpha spectrin (amino acids 965-1025) in presence of 100 mM 5-methyl uridine

Spectrin alpha chain, non-erythrocytic 1
MWexperimental 4 kDa
MWexpected 7 kDa
VPorod 10 nm3
log I(s) 1.05×10-3 1.05×10-4 1.05×10-5 1.05×10-6
Spectrin alpha chain, non-erythrocytic 1 small angle scattering data  s, nm-1
ln I(s)
Spectrin alpha chain, non-erythrocytic 1 Guinier plot ln 1.06×10-3 Rg: 1.3 nm 0 (1.3 nm)-2 s2
(sRg)2I(s)/I(0)
Spectrin alpha chain, non-erythrocytic 1 Kratky plot 1.104 0 √3 sRg
Dmax: 4.5 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from a solution of 10.00 mg/ml SPC-SH3 domain in 10 mM citric acid pH 3, 150 mM NaCl, 100 mM 5-methyl uridine was collected on the BL11-NCD-SWEET beam line at ALBA Synchrotron using a Pilatus 1M photon-counting detector. The beam at sample position was 0.566 mm (horizontal) × 0.420 mm (vertical), the incident photon flux was 1.08×10e12 photons/s at 12.4 keV (λ =0.0999 nm) and the sample to detector distance was 2.460 m. 2000 successive 0.1 second frames were collected and data were normalized to the intensity of the transmitted beam and radially averaged. The current buffer-subtracted scattering profile corresponds to the average of 208 collected frames. The absorbed dose by the sample was 18.10 kGy.

Sample temperature: 25°C. CAUTION! The scattering intensities do not have associated errors.

Spectrin alpha chain, non-erythrocytic 1 (SH3-SPC)
Mol. type   Protein
Organism   Gallus gallus
Olig. state   Unknown
Mon. MW   7.2 kDa
 
UniProt   P07751 (965-1025)
Sequence   FASTA