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.

SASDSU7 – Gramicidin Synthetase 1 (GrsA) W239S mutant in the holo form in the presence of 4 molar excess of L-Tyrosine

Gramicidin S synthase 1 (W239S)
MWexperimental 109 kDa
MWexpected 126 kDa
VPorod 181 nm3
log I(s) 4.35×101 4.35×100 4.35×10-1 4.35×10-2
Gramicidin S synthase 1 (W239S) small angle scattering data  s, nm-1
ln I(s)
Gramicidin S synthase 1 (W239S) Guinier plot ln 4.36×101 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
p(r)
Gramicidin S synthase 1 (W239S) pair distance distribution function Rg: 4.6 nm 0 Dmax: 17.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1

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

Synchrotron SAXS data from solutions of Gramicidin Synthetase 1 (GrsA) W239S mutant in the holo form in the presence of L-Tyrosine 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 holo form of GrsA-W239S exists in an ensemble of 2 states as determined by bilboMD in presence of L-Tyrosine. Individually, the compact form is present as 66% of the representative species with Dmax of 15.2 nm and Rg of 3.88 nm, while the elongated form was present as 34% of the ensemble with Dmax of 15.4 nm and Rg of 4.69 nm.

Gramicidin S synthase 1 (W239S) (GrsA-W239S)
Mol. type   Protein
Organism   Brevibacillus brevis
Olig. state   Monomer
Mon. MW   126.5 kDa
 
UniProt   P0C062 (1-1098)
Sequence   FASTA