SEC-SAXS characterization of the PPARγ2 AF-1 domain under varying pH and salt conditions

Mithun Karadi.

SASD2M8 – Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain (AF1) in 10 mM sodium phosphate, pH 8, containing 150 mM NaCl

Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain
MWexperimental 13 kDa
MWexpected 15 kDa
VPorod 17 nm3
log I(s) 5.77×10-1 5.77×10-2 5.77×10-3 5.77×10-4
Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain small angle scattering data  s, nm-1
ln I(s)
Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain Guinier plot ln 5.78×10-1 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain Kratky plot 1.104 0 √3 sRg
p(r)
Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain pair distance distribution function Rg: 3.1 nm 0 Dmax: 10.9 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain (AF1) in 10 mM sodium phosphate, pH 8, containing 150 mM NaCl in 10 mM sodium phosphate, 150 mM NaCl, pH 8 were collected on the ID7A1 BioSAXS / HP-Bio Beamline beam line at the Cornell High Energy Synchrotron Source (CHESS) storage ring (Ithaca, NY, USA) using a Eiger 4M detector at a sample-detector distance of 1.7 m and at a wavelength of λ = 0.110659 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). A 100 µL sample (10 mg/ml) was injected onto a Superdex 200 Increase 5/150 GL column and eluted isocratically at 0.30 mL/min at 4 °C, with 1 s SAXS exposures collected throughout the SEC run. The SEC-SAXS series was processed using BioXTAS RAW 2.3.0. Evolving factor analysis (EFA) identified a single significant scattering population corresponding to the main SEC elution peak, spanning frames 400–559. The resulting EFA-derived scattering profile was subsequently analyzed by Guinier and Bayesian indirect Fourier transform (BIFT) analyses.

Peroxisome Proliferator-Activated Receptor Gamma Activation Function-1 Domain (PPARγ AF-1 Domain)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   15.3 kDa
 
UniProt   P37231-1 (1-135)
Sequence   FASTA