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

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

SASDPF7 – Ribonuclease A SEC-SAXS data (EMBL-P12 bioSAXS beam line at DESY)

Ribonuclease pancreatic
MWexperimental 15 kDa
MWexpected 14 kDa
VPorod 16 nm3
log I(s) 1.00×10-2 1.00×10-3 1.00×10-4 1.00×10-5
Ribonuclease pancreatic small angle scattering data  s, nm-1
ln I(s)
Ribonuclease pancreatic Guinier plot ln 1.00×10-2 Rg: 1.5 nm 0 (1.5 nm)-2 s2
(sRg)2I(s)/I(0)
Ribonuclease pancreatic Kratky plot 1.104 0 3 sRg
p(r)
Ribonuclease pancreatic pair distance distribution function Rg: 1.5 nm 0 Dmax: 4.6 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Ribonuclease A in 50 mM HEPES, 150 mM KCl, 3% glycerol, pH 7.5 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 3 m and at a wavelength of λ = 0.124 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 75.00 μl sample at 8 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 75 Increase 10/300 column at 20°C. 30 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 (2400 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: https://www.sasbdb.org/project/1742/

Tags: benchmark
Ribonuclease pancreatic (RNaseA)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   13.7 kDa
 
UniProt   P61823 (27-150)
Sequence   FASTA