Structure of relaxase from Bacillus subtilis conjugative plasmid pLS20 reveals a new mechanism of DNA processing mediated by dimerization

Isidro Crespo.

SASDX82 – N-terminal domain of relaxase from Bacillus subtilis conjugative plasmid pLS20

Relaxase
MWexperimental 26 kDa
MWexpected 29 kDa
VPorod 48 nm3
log I(s) 1.91×100 1.91×10-1 1.91×10-2 1.91×10-3
Relaxase small angle scattering data  s, nm-1
ln I(s)
Relaxase Guinier plot ln 1.91×100 Rg: 2.3 nm 0 (2.3 nm)-2 s2
(sRg)2I(s)/I(0)
Relaxase Kratky plot 1.104 0 √3 sRg
p(r)
Relaxase pair distance distribution function Rg: 2.3 nm 0 Dmax: 7.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Relaxase PHENIX model
Synchrotron SAXS data from solutions of the N-terminal domain of relaxase in 20 mM Tris, 300 mM NaCl, 1 mM DTT, pH 8 were collected on the BL11-NCD beam line at ALBA (Cerdanyola del Vallès, Barcelona, Spain) using a ADSC Quantum 210r detector at a sample-detector distance of 2.6 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 25°C. 20 successive 0.100 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.

PLS20: https://doi.org/10.1093/nargab/lqab096

Relaxase
Mol. type   Protein
Organism   Bacillus subtilis subsp. natto
Olig. state   Monomer
Mon. MW   28.5 kDa
 
UniProt   E9RJ23 (1-232)
Sequence   FASTA