Combination of SAXS and Protein Painting Discloses the Three-Dimensional Organization of the Bacterial Cysteine Synthase Complex, a Potential Target for Enhancers of Antibiotic Action.

Rosa B, Marchetti M, Paredi G, Amenitsch H, Franko N, Benoni R, Giabbai B, De Marino MG, Mozzarelli A, Ronda L, Storici P, Campanini B, Bettati S, Int J Mol Sci 20(20) (2019) Europe PMC

SASDGW6 – Serine acetyltransferase

Serine acetyltransferase
MWexperimental 189 kDa
MWexpected 177 kDa
VPorod 280 nm3
log I(s) 1.91×101 1.91×100 1.91×10-1 1.91×10-2
Serine acetyltransferase small angle scattering data  s, nm-1
ln I(s)
Serine acetyltransferase Guinier plot ln 1.91×101 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Serine acetyltransferase Kratky plot 1.104 0 √3 sRg
p(r)
Serine acetyltransferase pair distance distribution function Rg: 3.9 nm 0 Dmax: 13 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Serine acetyltransferase DAMMIF model
Synchrotron SAXS data from solutions of serine acetyltransferase in 20 mM sodium phosphate, 85 mM NaCl, 2 mM EDTA, 10 mM 2-MCE, pH 7.5 were collected on the Austrian SAXS beamline 5.2L beam line at the ELETTRA - Sincrotrone Trieste storage ring (Trieste, Italy) using a Dectris / Pilatus3 1M detector at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.5 and 1.5 mg/ml were measured at 20°C. 12 successive 10 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Sample detector distance = UNKNOWN

Serine acetyltransferase (CysE)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Hexamer
Mon. MW   29.4 kDa
 
UniProt   P0A9D4 (2-273)
Sequence   FASTA