Solution characterization of the dynamic conjugative entry exclusion protein TraG

Bragagnolo N, Audette G, Structural Dynamics 9(6):064702 (2022) DOI

SASDQG6 – N-terminal truncation mutant of the periplasmic region of conjugal transfer mating pair stabilization protein TraG* from the R100 plasmid

Conjugal transfer mating pair stabilization protein TraG
MWI(0) 53 kDa
MWexpected 52 kDa
VPorod 130 nm3
log I(s) 1.51×10-2 1.51×10-3 1.51×10-4 1.51×10-5
Conjugal transfer mating pair stabilization protein TraG small angle scattering data  s, nm-1
ln I(s)
Conjugal transfer mating pair stabilization protein TraG Guinier plot ln 1.51×10-2 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Conjugal transfer mating pair stabilization protein TraG Kratky plot 1.104 0 √3 sRg
p(r)
Conjugal transfer mating pair stabilization protein TraG pair distance distribution function Rg: 4.4 nm 0 Dmax: 17.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Conjugal transfer mating pair stabilization protein TraG DAMMIF model
Synchrotron SAXS data from solutions of a N-terminal truncation mutant of TraG* in 20 mM HEPES, 100 mM NaCl, 5% glycerol, pH 7 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS; Argonne National Laboratory USA) using a Eiger2 XE 9M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.1033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 300.00 μl sample at 4.8 mg/ml was injected at a 0.6 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 2594 successive 0.500 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 sample underwent sequential multi-angle laser light scattering analysis using a a Wyatt DAWN Heleos MALS system.

Conjugal transfer mating pair stabilization protein TraG (deltaTraG*)
Mol. type   Protein
Organism   Shigella flexneri 4c
Olig. state   Monomer
Mon. MW   52.0 kDa
 
UniProt   A0A0C5PJM1 (497-940)
Sequence   FASTA