A unique hetero-hexadecameric architecture displayed by the Escherichia coli O157 PaaA2-ParE2 antitoxin-toxin complex.

Sterckx YG, Jové T, Shkumatov AV, Garcia-Pino A, Geerts L, De Kerpel M, Lah J, De Greve H, Van Melderen L, Loris R, J Mol Biol 428(8):1589-603 (2016) Europe PMC

SASDC84 – parDE-like toxin-antitoxin module, EcPaaA2_13-63-HisEcParE2 construct

Plasmid stabilization protein ParE
Uncharacterized protein
MWexperimental 21 kDa
MWexpected 19 kDa
VPorod 36 nm3
log I(s) 2.13×10-2 2.13×10-3 2.13×10-4 2.13×10-5
Plasmid stabilization protein ParE Uncharacterized protein small angle scattering data  s, nm-1
ln I(s)
Plasmid stabilization protein ParE Uncharacterized protein Guinier plot ln 2.14×10-2 Rg: 2.2 nm 0 (2.2 nm)-2 s2
(sRg)2I(s)/I(0)
Plasmid stabilization protein ParE Uncharacterized protein Kratky plot 1.104 0 √3 sRg
p(r)
Plasmid stabilization protein ParE Uncharacterized protein pair distance distribution function Rg: 2.2 nm 0 Dmax: 9.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
parDE-like toxin-antitoxin module, EcPaaA2_13-63-HisEcParE2 construct Rg histogram Rg, nm
Plasmid stabilization protein ParE Uncharacterized protein EOM/RANCH model
Plasmid stabilization protein ParE Uncharacterized protein EOM/RANCH model
Plasmid stabilization protein ParE Uncharacterized protein EOM/RANCH model
Synchrotron SAXS data from EcPaaA2_13-63-EcParE2 were collected at the SWING beamline (SOLEIL synchrotron, Gif-sur-Yvette, France) in HPLC mode using a CCD AVIEX detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.103 nm (s = 4π sin θ/λ, where 2θ is the scattering angle). Sample was loaded onto a Shodex KW402.5-4F, which was pre-equilibrated with running buffer (50 mM Tris-HCl, 500 mM NaCl, pH 7.5) for at least one column volume. The flow rate was set to 0.2 ml/min. During each HPLC run, 250 frames were collected after 15 min (0.62 CV) using 750 ms per frame and 1500 ms dead time. After radial averaging and buffer subtraction of the data frames using FOXTROT, the scattering curves were processed and analyzed with DATASW.

Concentration min = UNKNOWN

Plasmid stabilization protein ParE (ParE2)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   12.6 kDa
 
UniProt   A0A0D7C2L1 (2-92)
Sequence   FASTA
 
Uncharacterized protein (PaaA2)
Mol. type   Protein
Organism   Escherichia coli O157:H7
Olig. state   Monomer
Mon. MW   6.2 kDa
 
UniProt   A0A0F6F6Q9 (13-63)
Sequence   FASTA