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

SASDBB9 – GDP bound form of full length ObgE from E.coli (ObgE_FL with GDP)

GTPase ObgE/CgtA
MWexperimental 45 kDa
MWexpected 44 kDa
VPorod 94673 nm3
log I(s) 1.53×10-2 1.53×10-3 1.53×10-4 1.53×10-5
GTPase ObgE/CgtA small angle scattering data  s, nm-1
ln I(s)
GTPase ObgE/CgtA Guinier plot ln 1.54×10-2 Rg: 3.5 nm 0 (3.5 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: 3.6 nm 0 Dmax: 16.0 nm

Data validation

Fits and models

log I(s)
 s, nm-1
GTPase ObgE/CgtA DAMMIN model
Synchrotron SAXS data from solutions of the GDP bound form of full length ObgE from E.coli (ObgE_FL 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 SWING beam line at the SOLEIL Synchrotron (Saint-Aubin, France) using a CCD AVIEX detector at a sample-detector distance of 2.0 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). Data collection was performed at at 10°C. 255 successive 2.8 second frames were collected through an HPLC-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 and the different curves were scaled for protein concentration, then averaged to produce the final SAXS profile displayed in this entry. The sample protein includes a C-terminal Strep-tag; frames corresponding to the peak in HPLC-SAXS run were averaged using DATAVER v0.2 (r3709).

GTPase ObgE/CgtA (ObgE_FL)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   44.4 kDa
 
UniProt   P42641 (1-390)
Sequence   FASTA