Resting state of GrsA is a combination of two constitutively inactive isoforms that manifest long-range allosteric effects in presence of aminoacid and ATP

Raktim Roy.

SASDSR7 – Gramicidin Synthetase 1 (GrsA) W239S mutant in the apo form without any substrates

Gramicidin S synthase 1 (W239S)
MWexperimental 113 kDa
MWexpected 127 kDa
VPorod 187 nm3
log I(s) 1.12×102 1.12×101 1.12×100 1.12×10-1
Gramicidin S synthase 1 (W239S) small angle scattering data  s, nm-1
ln I(s)
Gramicidin S synthase 1 (W239S) Guinier plot ln 1.13×102 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Gramicidin S synthase 1 (W239S) Kratky plot 1.104 0 3 sRg
Dmax: 15 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Gramicidin S synthase 1 (W239S) BILBOMD model
Gramicidin S synthase 1 (W239S) BILBOMD model

log I(s)
 s, nm-1

Synchrotron SAXS data from solutions of Gramicidin Synthetase 1 (GrsA) W239S mutant in the apo form in 20 mM Tris pH 7.5, 100 mM NaCl, 10 mM MgCl2, 2% glycerol, were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS; Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2.1 m and at a wavelength of λ = 0.1127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 7.9°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The apo form of GrsA-W239S exists in an ensemble of 2 states as determined by bilboMD. Individually, the compact form is present as 51% of the representative species with Dmax of 14.6 nmand Rg of 3.93 nm, while the elongated form was present as 49% of the ensemble with Dmax of 15.4 nm and Rg of 4.69 nm. The atomic models displayed in this entry represent the compact form (top) and elongated form (bottom).

Gramicidin S synthase 1 (W239S) (GrsA-W239S)
Mol. type   Protein
Organism   Aneurinibacillus migulanus
Olig. state   Monomer
Mon. MW   126.5 kDa
 
UniProt   P0C061 (1-1098)
Sequence   FASTA