A structural model of PpoA derived from SAXS-analysis-implications for substrate conversion.

Koch C, Tria G, Fielding AJ, Brodhun F, Valerius O, Feussner K, Braus GH, Svergun DI, Bennati M, Feussner I, Biochim Biophys Acta 1831(9):1449-57 (2013) Europe PMC

SASDA45 – PpoA wild type enzyme

Psi-producing oxygenase A
MWexperimental 330 kDa
MWexpected 362 kDa
VPorod 540 nm3
log I(s) 2.68×101 2.68×100 2.68×10-1 2.68×10-2
Psi-producing oxygenase A small angle scattering data  s, nm-1
ln I(s)
Psi-producing oxygenase A Guinier plot ln 2.68×101 Rg: 5.4 nm 0 (5.4 nm)-2 s2
(sRg)2I(s)/I(0)
Psi-producing oxygenase A Kratky plot 1.104 0 √3 sRg
p(r)
Psi-producing oxygenase A pair distance distribution function Rg: 5.4 nm 0 Dmax: 16.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Psi-producing oxygenase A SASREF model
Synchrotron SAXS data from solutions of PpoA wild type enzyme in 20 Mm HEPES, pH 7.5 were collected on the EMBL X33 beam line at the DORIS III, DESY storage ring (Hamburg, Germany) using a Pilatus 1M-W detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.15 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.4 and 26 mg/ml were measured at 10°C. Eight successive 120 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.

Tags: X33
Psi-producing oxygenase A (PpoA)
Mol. type   Protein
Organism   Aspergillus nidulans
Olig. state   Trimer
Mon. MW   120.8 kDa
 
UniProt   Q6RET3
Sequence   FASTA