Small-Angle X-ray Scattering data (benchmarking/consensus): EMBL-P12 SAXS beam line, DESY

Clement Blanchet, Melissa Graewert, Cy M Jeffries, Dmitri Svergun.

SASDPJ7 – Urate Oxidase SEC-WAXS data (EMBL-P12 bioSAXS beam line at DESY)

Urate Oxidase (Uricase) from Aspergillus flavus
MWexperimental 135 kDa
MWexpected 137 kDa
VPorod 214 nm3
log I(s) 7.85×10-2 7.85×10-3 7.85×10-4 7.85×10-5
Urate Oxidase (Uricase) from Aspergillus flavus small angle scattering data  s, nm-1
ln I(s)
Urate Oxidase (Uricase) from Aspergillus flavus Guinier plot ln 7.85×10-2 Rg: 3.2 nm 0 (3.2 nm)-2 s2
Urate Oxidase (Uricase) from Aspergillus flavus Kratky plot 1.104 0 3 sRg
Urate Oxidase (Uricase) from Aspergillus flavus pair distance distribution function Rg: 3.2 nm 0 Dmax: 9 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Urate Oxidase in 100 mM Tris, 150 mM NaCl, 1 % v/v glycerol, pH 8 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.062 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 82.00 μl sample at 5.9 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 21 successive 1 second frames were collected through the main SEC elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The quoted experimental MW was determined from MALLS/RI data. The full entry .zip archive contains MW estimates from MALLS/UV and from I(0) and concentration in addition to the relative- and absolute-scaled unsubtracted SEC-SAXS data frames collected through the entire SEC-elution (2100 data frames), the CHROMIXS Rg-correlation through the SEC-elution peak, and the selected unsubtracted sample and buffer frames used to generate the profile displayed in this entry. These data form part of the 'Consensus and benchmarking SAS data project' by Trewhella et al., 2022:

Tags: benchmark
Urate Oxidase (Uricase) from Aspergillus flavus (Urate Oxidase)
Mol. type   Protein
Organism   Aspergillus flavus
Olig. state   Tetramer
Mon. MW   34.2 kDa
UniProt   Q00511 (1-302)
Sequence   FASTA