Structural and Biochemical Analysis of Escherichia coli ObgE, a Central Regulator of Bacterial Persistence

Gkekas S, Singh R, Shkumatov A, Messens J, Fauvart M, Verstraeten N, Michiels J, Versées W, Journal of Biological Chemistry 2017 Feb 21 :jbc.M116.761809 DOI

SASDBS6 – Apo form of full length ObgE from E.coli (ObgE_FL)

GTPase ObgE/CgtA
MWexperimental 55.4 kDa
MWexpected 44 kDa
VPorod 102 nm3
log I(s) 3.83×10-2 3.83×10-3 3.83×10-4 3.83×10-5
GTPase ObgE/CgtA small angle scattering data  s, nm-1
ln I(s)
GTPase ObgE/CgtA Guinier plot ln 3.83×10-2 Rg: 3.7 nm 0 (3.7 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: 4.0 nm 0 Dmax: 18.1 nm
log I(s)
 s, nm-1
Apo form of full length ObgE from E.coli (ObgE_FL) Rg histogram Rg, nm
GTPase ObgE/CgtA RANCH/EOM model
GTPase ObgE/CgtA RANCH/EOM model
GTPase ObgE/CgtA RANCH/EOM model
GTPase ObgE/CgtA RANCH/EOM model
GTPase ObgE/CgtA RANCH/EOM model
log I(s)
 s, nm-1
GTPase ObgE/CgtA DAMMIN model
Synchrotron SAXS data from solutions of the Apo form of full length ObgE from E.coli (ObgE_FL) 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.

SAXS-based ensemble modelling of ObgE_FL taking into account the flexibility of the C-terminal domain. 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 (strain K12)
Olig. state   Monomer
Mon. MW   44.4 kDa
 
UniProt   P42641 (1-390)
Sequence   FASTA