Quantitative live-cell imaging and 3D modeling reveal critical functional features in the cytosolic complex of phagocyte NADPH oxidase.

Ziegler CS, Bouchab L, Tramier M, Durand D, Fieschi F, Dupré-Crochet S, Mérola F, Nüße O, Erard M, J Biol Chem (2019) Europe PMC

SASDEJ3 – Truncated neutophil cytosol factor 1, p47phox [1-342]

Neutophil cytosol factor 1
MWI(0) 38 kDa
MWexpected 40 kDa
VPorod 58 nm3
log I(s) 2.87×10-2 2.87×10-3 2.87×10-4 2.87×10-5
Neutophil cytosol factor 1 small angle scattering data  s, nm-1
ln I(s)
Neutophil cytosol factor 1 Guinier plot ln 2.87×10-2 Rg: 2.6 nm 0 (2.6 nm)-2 s2
Neutophil cytosol factor 1 Kratky plot 1.104 0 3 sRg
Neutophil cytosol factor 1 pair distance distribution function Rg: 2.6 nm 0 Dmax: 10 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Neutophil cytosol factor 1 OTHER model

SAXS data from solutions of p47phox [1-342] in 50 mM HEPES, 100 mM NaCl, 1 mM EDTA, 2 mM DTT, 5% glycerol, pH 7.5, were collected on a Bruker Nanostar instrument at the IBBMC (Orsay, France) using a MikroGap VÅNTEC-2000 detector at a sample-detector distance of 0.7 m and at a wavelength of λ = 0.154 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Samples of were studied at different concentrations ranging from 0.9 to 3.6 mg/ml measured at 6°C. 11 successive 3600 second frames were collected. SAXS data were normalized to the intensity of the incident beam, averaged and background subtracted using the program package PRIMUS. Intensities were put on an absolute scale (cm-1) using water scattering. The final pattern used for fitting was obtained by extrapolating to infinite dilution of the set of curves recorded at the different concentrations. Molecular mass was obtained using the bayesian inference approach proposed in PRIMUS/qt, which combines four concentration independent MM estimators: M=39.4 kDa within the credibility interval [36.5-39.8].

Neutophil cytosol factor 1 (p47phox)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   40.4 kDa
UniProt   P14598 (1-342)
Sequence   FASTA