Inline small‐angle X‐ray scattering‐coupled chromatography under extreme hydrostatic pressure

Miller R, Cummings C, Huang Q, Ando N, Gillilan R, Protein Science 31(12) (2022) DOI

SASDP57 – Glucose Isomerase at a neutral pH and 100 MPa (1000 atm) of pressure

Xylose isomerase
MWexperimental 170 kDa
MWexpected 173 kDa
VPorod 267 nm3
log I(s) 2.52×10-2 2.52×10-3 2.52×10-4 2.52×10-5
Xylose isomerase small angle scattering data  s, nm-1
ln I(s)
Xylose isomerase Guinier plot ln 2.53×10-2 Rg: 3.5 nm 0 (3.5 nm)-2 s2
Xylose isomerase Kratky plot 1.104 0 3 sRg
Dmax: 10.6 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of glucose isomerase in 25 mM HEPES, 150 mM NaCl, and 3% v/v glycerol, pH 7 were collected on the ID7A1 BioSAXS / HP-Bio Beamline beam line at the Cornell High Energy Synchrotron Source (CHESS; Ithaca, NY, USA) using a Eiger 4M detector at a sample-detector distance of 1.6 m and at a wavelength of λ = 0.089 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 sample at 17.9 mg/ml was injected onto a column at 23°C. 82 successive 2 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.

SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

Xylose isomerase
Mol. type   Protein
Organism   Streptomyces rubiginosus
Olig. state   Tetramer
Mon. MW   43.2 kDa
UniProt   P24300 (1-380)
Sequence   FASTA