The Subnanometer Resolution Structure of the Glutamate Synthase 1.2-MDa Hexamer by Cryoelectron Microscopy and Its Oligomerization Behavior in Solution

Cottevieille M, Larquet E, Jonic S, Petoukhov M, Caprini G, Paravisi S, Svergun D, Vanoni M, Boisset N, Journal of Biological Chemistry 283(13):8237-8249 (2008) DOI

SASDLZ6 – Hexamer - monomer equilibrium of Glutamate Synthase

Glutamate synthase [NADPH] large chain
Glutamate synthase [NADPH] small chain
MWexperimental 1419 kDa
MWexpected 1264 kDa
log I(s) 5.48×103 5.48×102 5.48×101 5.48×100
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain small angle scattering data  s, nm-1
ln I(s)
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain Guinier plot ln 5.49×103 Rg: 7.7 nm 0 (7.7 nm)-2 s2
(sRg)2I(s)/I(0)
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain Kratky plot 1.104 0 √3 sRg
p(r)
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain pair distance distribution function Rg: 7.5 nm 0 Dmax: 22.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain PDB (PROTEIN DATA BANK) model
Glutamate synthase [NADPH] large chain Glutamate synthase [NADPH] small chain PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of Hexamer - monomer equilibrium of Glutamate Synthase in 25 mM Hepes/KOH, 1 mM EDTA, 1 mM dithiothreitol, pH 7.5 were collected on the EMBL X33 beam line at the DORIS III, DESY storage ring (Hamburg, Germany) using a MAR 345 Image Plate 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). One solute concentration of 0.80 mg/ml was measured. Two successive 60 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.

Cell temperature = UNKNOWN. Storage temperature = UNKNOWN

Tags: X33
Glutamate synthase [NADPH] large chain (Glts A subunit)
Mol. type   Protein
Organism   Azospirillum brasilense
Olig. state   Hexamer
Mon. MW   161.4 kDa
 
UniProt   Q05755 (1-1508)
Sequence   FASTA
 
Glutamate synthase [NADPH] small chain (Glts B subunit)
Mol. type   Protein
Organism   Azospirillum brasilense
Olig. state   Hexamer
Mon. MW   49.3 kDa
 
UniProt   Q05756 (27-482)
Sequence   FASTA
 
PDB ID   2VDC
 
PDB ID   2VDC