Long-range structural defects by pathogenic mutations in most severe glucose-6-phosphate dehydrogenase deficiency

Horikoshi N, Hwang S, Gati C, Matsui T, Castillo-Orellana C, Raub A, Garcia A, Jabbarpour F, Batyuk A, Broweleit J, Xiang X, Chiang A, Broweleit R, Vöhringer-Martinez E, Mochly-Rosen D, Wakatsuki S, Proceedings of the National Academy of Sciences 118(4) (2021) DOI

SASDJ45 – Glucose-6-phosphate dehydrogenase P396L mutant

Glucose-6-phosphate 1-dehydrogenase P396L
MWexperimental 119 kDa
MWexpected 119 kDa
VPorod 178 nm3
log I(s) 1.37×10-1 1.37×10-2 1.37×10-3 1.37×10-4
Glucose-6-phosphate 1-dehydrogenase P396L small angle scattering data  s, nm-1
ln I(s)
Glucose-6-phosphate 1-dehydrogenase P396L Guinier plot ln 1.37×10-1 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
Glucose-6-phosphate 1-dehydrogenase P396L Kratky plot 1.104 0 √3 sRg
p(r)
Glucose-6-phosphate 1-dehydrogenase P396L pair distance distribution function Rg: 3.7 nm 0 Dmax: 13 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Glucose-6-phosphate 1-dehydrogenase P396L CORAL model
Synchrotron SAXS data from solutions of Glucose-6-phosphate dehydrogenase P396L mutant in 20 mM Tris, 150 mM NaCl, pH 8 were collected on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSRL) storage ring (Menlo Park, CA, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 1.7 m and at a wavelength of λ = 0.112756 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 100.00 μl sample at 5 mg/ml was injected at a 0.05 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 25°C. Five successive 1 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.

Glucose-6-phosphate 1-dehydrogenase P396L (G6PD P396L mutant)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   59.6 kDa
 
UniProt   P11413 (1-515)
Sequence   FASTA