SAXS studies of KAI2 catalytic mutants

Sabrina Davies.

SASDS28 – Karrikin insensitive 2, KAI2 protein (aggregate peak)

Probable esterase KAI2 (KARRIKIN INSENSITIVE 2)
MWexperimental 184 kDa
MWexpected 30 kDa
VPorod 283 nm3
log I(s) 1.12×10-2 1.12×10-3 1.12×10-4 1.12×10-5
Probable esterase KAI2 (KARRIKIN INSENSITIVE 2) small angle scattering data  s, nm-1
ln I(s)
Probable esterase KAI2 (KARRIKIN INSENSITIVE 2) Guinier plot ln 1.13×10-2 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Probable esterase KAI2 (KARRIKIN INSENSITIVE 2) Kratky plot 1.104 0 3 sRg
p(r)
Probable esterase KAI2 (KARRIKIN INSENSITIVE 2) pair distance distribution function Rg: 4.4 nm 0 Dmax: 15.8 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Karrikin insensitive 2, KAI2 protein (aggregate peak) in 20 mM HEPES, 150 mM imidazole, 150 mM NaCl, 10% (v/v) glycerol, pH 7.5 were collected on the SAXS/WAXS beam line at the Australian Synchrotron storage ring (Melbourne, Australia) using a Pilatus3 S 2M detector at a sample-detector distance of 2.5 m and at a wavelength of λ = 0.10332 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 8.1 mg/ml was injected at a 0.40 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 24°C. 559 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 experimental MW value was determined using the volume of correlation, Vc, method.

Probable esterase KAI2 (KARRIKIN INSENSITIVE 2) (KAI2)
Mol. type   Protein
Organism   Arabidopsis thaliana
Olig. state   Monomer
Mon. MW   29.8 kDa
 
UniProt   Q9SZU7
Sequence   FASTA