Solution characterization of the dynamic conjugative entry exclusion protein TraG

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

SASDQH6 – Periplasmic region of the conjugal transfer mating pair stabilization protein TraG* from the R100 plasmid

Conjugal transfer mating pair stabilization protein TraG
MWI(0) 198 kDa
MWexpected 57 kDa
log I(s) 7.13×10-1 7.13×10-2 7.13×10-3 7.13×10-4
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 7.14×10-1 Rg: 5.8 nm 0 (5.8 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: 9.1 nm 0 Dmax: 45 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 the periplasmic region of TraG* in 20 mM HEPES, 100 mM NaCl, 5% glycerol, 0.05% NP40, 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 5 mg/ml was injected at a 0.6 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 3025 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. Protein aggregation was present in the sample resulting in data that may be inaccurate.

Conjugal transfer mating pair stabilization protein TraG (TraG*)
Mol. type   Protein
Organism   Shigella flexneri 4c
Olig. state   Monomer
Mon. MW   56.7 kDa
 
UniProt   A0A0C5PJM1 (452-940)
Sequence   FASTA