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

SASDBU6 – Apo form of the C-terminal deletion mutant of ObgE from E.coli (ObgE_340)

GTPase ObgE/CgtA
MWexperimental 41 kDa
MWexpected 39 kDa
VPorod 70 nm3
log I(s) 3.68×10-2 3.68×10-3 3.68×10-4 3.68×10-5
GTPase ObgE/CgtA small angle scattering data  s, nm-1
ln I(s)
GTPase ObgE/CgtA Guinier plot ln 3.68×10-2 Rg: 3.1 nm 0 (3.1 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: 11.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Apo form of the C-terminal deletion mutant of ObgE from E.coli (ObgE_340) Rg histogram Rg, nm
GTPase ObgE/CgtA EOM/RANCH model
GTPase ObgE/CgtA EOM/RANCH model
GTPase ObgE/CgtA EOM/RANCH model
GTPase ObgE/CgtA EOM/RANCH model

log I(s)
 s, nm-1
GTPase ObgE/CgtA DAMMIN model

Synchrotron SAXS data from solutions of the Apo form of the C-terminal deletion mutant of ObgE from E.coli (ObgE_340) in 20 mM HEPES, 300 mM NaCl, 250 mM imidazole, 5 mM MgCl2, 2 mM DTT, 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 an N-terminal poly-histidine tag; frames corresponding to the peak in HPLC-SAXS run were averaged using DATAVER v0.2 (r3709)

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