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