Alternative dimerization is required for activity and inhibition of the HEPN ribonuclease RnlA

Garcia-Rodriguez G, Charlier D, Wilmaerts D, Michiels J, Loris R, Nucleic Acids Research 49(12):7164-7178 (2021) DOI

SASDHW7 – RnlA-RnlB toxin-antitoxin complex (mRNA endoribonuclease toxin LS : Antitoxin RnlB)

mRNA endoribonuclease toxin LS
Antitoxin RnlB (C-terminal His-tagged)
MWexperimental 100 kDa
MWexpected 95 kDa
VPorod 161 nm3
log I(s) 5.79×101 5.79×100 5.79×10-1 5.79×10-2
mRNA endoribonuclease toxin LS Antitoxin RnlB (C-terminal His-tagged) small angle scattering data  s, nm-1
ln I(s)
mRNA endoribonuclease toxin LS Antitoxin RnlB (C-terminal His-tagged) Guinier plot ln 5.79×101 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
mRNA endoribonuclease toxin LS Antitoxin RnlB (C-terminal His-tagged) Kratky plot 1.104 0 3 sRg
p(r)
mRNA endoribonuclease toxin LS Antitoxin RnlB (C-terminal His-tagged) pair distance distribution function Rg: 3.4 nm 0 Dmax: 10.0 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of RnlA-RnlB toxin-antitoxin complex (mRNA endoribonuclease toxin LS : Antitoxin RnlB) in 20 mM Tris, 150 mM NaCl, 1 mM TCEP, 5% glycerol, pH 8 were collected on the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.099 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 50.00 μl sample at 5.9 mg/ml was injected at a 0.20 ml/min flow rate onto a Shodex KW402.5-4F column at 20°C. 900 successive 2 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

mRNA endoribonuclease toxin LS (RnlA)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Dimer
Mon. MW   40.1 kDa
 
UniProt   P52129 (1-357)
Sequence   FASTA
 
Antitoxin RnlB (C-terminal His-tagged) (RnlB)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   14.9 kDa
 
UniProt   P52130 (1-123)
Sequence   FASTA