Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument.

Bucciarelli S, Midtgaard SR, Nors Pedersen M, Skou S, Arleth L, Vestergaard B, J Appl Crystallogr 51(Pt 6):1623-1632 (2018) Europe PMC

SASDEE5 – Bovine serum albumin from synchrotron SAXS

Bovine serum albumin
MWexperimental 66 kDa
MWexpected 66 kDa
VPorod 85 nm3
log I(s) 4.50×10-2 4.50×10-3 4.50×10-4 4.50×10-5
Bovine serum albumin small angle scattering data  s, nm-1
ln I(s)
Bovine serum albumin Guinier plot ln 4.50×10-2 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Bovine serum albumin Kratky plot 1.104 0 √3 sRg
p(r)
Bovine serum albumin pair distance distribution function Rg: 2.8 nm 0 Dmax: 9.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Bovine serum albumin PDB (PROTEIN DATA BANK) model
log I(s)
 s, nm-1
Bovine serum albumin DAMFILT model
Synchrotron SAXS data from solutions of Bovine serum albumin in 50 mM HEPES, pH 7.5 were collected on the EMBL P12 beam line at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.30 mg/ml was measured at 20°C. 20 successive 0.050 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.

The data have been binned logarithmically and scaled to overlap with static in-house SAXS BSA data (SASDEB5).

Bovine serum albumin (BSA)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   66.4 kDa
 
UniProt   P02769 (25-607)
Sequence   FASTA
 
PDB ID   4F52