Automated pipeline for purification, biophysical and x-ray analysis of biomacromolecular solutions.

Graewert MA, Franke D, Jeffries CM, Blanchet CE, Ruskule D, Kuhle K, Flieger A, Schäfer B, Tartsch B, Meijers R, Svergun DI, Sci Rep 5:10734 (2015) Europe PMC

SASDA97 – PlaB

PlaB
MWI(0) 225 kDa
MWexpected 220 kDa
VPorod 270 nm3
log I(s) 1.38×103 1.38×102 1.38×101 1.38×100
PlaB small angle scattering data  s, nm-1
ln I(s)
PlaB Guinier plot ln 1.38×103 Rg: 4.0 nm 0 (4.0 nm)-2 s2
(sRg)2I(s)/I(0)
PlaB Kratky plot 1.104 0 √3 sRg
p(r)
PlaB pair distance distribution function Rg: 3.9 nm 0 Dmax: 10.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
PlaB DAMMIN model
Synchrotron SAXS data from solutions of PlaB in 100 mM Tris 100 mM Nacl, pH 7.5 were collected on the EMBL P12 beam line at the PETRA III storage ring (DESY; Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.12 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.1 and 4.5 mg/ml were measured . 160 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Cell temperature = UNKNOWN. Storage temperature = UNKNOWN

PlaB
Mol. type   Protein
Organism   Legionella pneumophila
Olig. state   Tetramer
Mon. MW   55 kDa