Structural and biochemical analysis of Escherichia coli ObgE, a central regulator of bacterial persistence.

Gkekas S, Singh RK, Shkumatov AV, Messens J, Fauvart M, Verstraeten N, Michiels J, Versées W, J Biol Chem 292(14):5871-5883 (2017) Europe PMC

SASDBC9 – GDP bound form of C-terminal deletion mutant of ObgE from E.coli (ObgE_340 with GDP)

GTPase ObgE/CgtA
MWexperimental 39 kDa
MWexpected 39 kDa
VPorod 70673 nm3
log I(s) 1.01×100 1.01×10-1 1.01×10-2 1.01×10-3
GTPase ObgE/CgtA small angle scattering data  s, nm-1
ln I(s)
GTPase ObgE/CgtA Guinier plot ln 1.01×100 Rg: 3.0 nm 0 (3.0 nm)-2 s2
(sRg)2I(s)/I(0)
GTPase ObgE/CgtA Kratky plot 1.104 0 √3 sRg
p(r)
GTPase ObgE/CgtA pair distance distribution function Rg: 3.2 nm 0 Dmax: 11 nm

Data validation

Fits and models

log I(s)
 s, nm-1
GTPase ObgE/CgtA DAMMIN model
SAXS data collected from solutions of the GDP bound form of the C-terminal deletion mutant of ObgE from E.coli (ObgE_340 with GDP) in 20 mM HEPES, 300 mM NaCl, 250 mM imidazole, 5 mM MgCl2, 2 mM DTT, 400 µM GDP, pH 7.5, were collected at the Rigaku BioSAXS-2000 instrument using a CCD detector at a sample-detector distance of 1.0 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). Data collection was performed at at 10°C at four different concentrations of 1, 2, 4 and 8 mg/ml with 30 min exposure each intermittent by reference buffer. Rigaku SAXSLab was used for data reduction. The sample protein includes an N-terminal poly-histidine tag. Data was merged using PRIMUS from ATSAS package.

GTPase ObgE/CgtA (ObgE_340)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   38.9 kDa
 
UniProt   P42641 (1-340)
Sequence   FASTA