Structural Basis for Membrane Targeting and Secretion of Legionella SidE Ubiquitin Ligases

Ahmed Mohammed.

SASDUW5 – C-terminal domain of Ubiquitinating/deubiquitinating enzyme SdeA (SdeA_CTD)

Ubiquitinating/deubiquitinating enzyme SdeA
MWexperimental 40 kDa
MWexpected 39 kDa
VPorod 62 nm3
log I(s) 2.09×104 2.09×103 2.09×102 2.09×101
Ubiquitinating/deubiquitinating enzyme SdeA small angle scattering data  s, nm-1
ln I(s)
Ubiquitinating/deubiquitinating enzyme SdeA Guinier plot ln 2.10×104 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Ubiquitinating/deubiquitinating enzyme SdeA Kratky plot 1.104 0 √3 sRg
p(r)
Ubiquitinating/deubiquitinating enzyme SdeA pair distance distribution function Rg: 4.2 nm 0 Dmax: 15.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Ubiquitinating/deubiquitinating enzyme SdeA MONSA model
log I(s)
 s, nm-1
Ubiquitinating/deubiquitinating enzyme SdeA SREFLEX model
Synchrotron SAXS data from solutions of SdeA_CTD in 25 mM Tris-HCl, 200 mM NaCl, 5 mM DTT, pH 7.5 were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 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 100.00 μl sample at 16.5 mg/ml was injected at a 0.70 ml/min flow rate onto a GE Superdex 75 Increase 10/300 column at 25°C. 36 successive 1 second frames were collected through the main SEC elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Ubiquitinating/deubiquitinating enzyme SdeA
Mol. type   Protein
Organism   Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)
Olig. state   Monomer
Mon. MW   39.4 kDa
 
UniProt   Q5ZTK4 (1158-1499)
Sequence   FASTA