Isolation and characterization of monomeric human RAD51: a novel tool for investigating homologous recombination in cancer

Rinaldi F, Schipani F, Balboni B, Catalano F, Marotta R, Myers S, Previtali V, Veronesi M, Scietti L, Cecatiello V, Pasqualato S, Ortega J, Girotto S, Cavalli A, Angewandte Chemie International Edition (2023) DOI

SASDQ97 – Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E]

DNA repair protein RAD51 homolog 1 (F86E A89E)
MWexperimental 41 kDa
MWexpected 37 kDa
VPorod 61 nm3
log I(s) 4.47×10-2 4.47×10-3 4.47×10-4 4.47×10-5
DNA repair protein RAD51 homolog 1 (F86E A89E) small angle scattering data  s, nm-1
ln I(s)
DNA repair protein RAD51 homolog 1 (F86E A89E) Guinier plot ln 4.48×10-2 Rg: 3.0 nm 0 (3.0 nm)-2 s2
(sRg)2I(s)/I(0)
DNA repair protein RAD51 homolog 1 (F86E A89E) Kratky plot 1.104 0 √3 sRg
p(r)
DNA repair protein RAD51 homolog 1 (F86E A89E) pair distance distribution function Rg: 3.2 nm 0 Dmax: 14.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E] Rg histogram Rg, nm
DNA repair protein RAD51 homolog 1 (F86E A89E) EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) EOM/RANCH model
log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E A89E) MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) MULTIFOXS model
log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E A89E) ALPHAFOLD model
Synchrotron SAXS data from solutions of the DNA repair protein RAD51 homolog 1, double mutant [F86E, A89E], in 20 mM HEPES, 200 mM Na2SO4, 5% glycerol, 0.1 mM EDTA, pH 8 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.09537 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 100.00 μl sample at 8.8 mg/ml was injected at a 0.07 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 15°C. 599 successive 3 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.

DNA repair protein RAD51 homolog 1 (F86E A89E) (RAD51 F86E A89E)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   37.1 kDa
 
UniProt   Q06609 (1-339)
Sequence   FASTA