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

SASDQT9 – Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E] in complex with fourth BRC repeat (BRC4)

DNA repair protein RAD51 homolog 1 (F86E A89E)
Breast cancer type 2 susceptibility protein
MWexperimental 42 kDa
MWexpected 42 kDa
VPorod 70 nm3
log I(s) 3.30×10-2 3.30×10-3 3.30×10-4 3.30×10-5
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein small angle scattering data  s, nm-1
ln I(s)
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein Guinier plot ln 3.30×10-2 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein Kratky plot 1.104 0 √3 sRg
p(r)
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein pair distance distribution function Rg: 3.1 nm 0 Dmax: 15.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E] in complex with fourth BRC repeat (BRC4) Rg histogram Rg, nm
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein EOM/RANCH model
log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein ALPHAFOLD model
log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein MULTIFOXS model
DNA repair protein RAD51 homolog 1 (F86E A89E) Breast cancer type 2 susceptibility protein MULTIFOXS model
Synchrotron SAXS data from solutions of DNA repair protein RAD51 homolog 1 [F86E, A89E] in complex with the fourth BRC repeat of BRC4 in 20 mM HEPES, 50 mM Na2SO4, 2% v/v sucrose, 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 = 0.09464 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 7 mg/ml was injected at a 0.07 ml/min flow rate onto a GE Superose 6 Increase 3.2/300 column at 15°C. 29 successive frames were collected through the sample elution peak (from a total of 599 frames). The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Experimental molecular weight has been calculated through volume of correlation (Vc).

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
 
Breast cancer type 2 susceptibility protein (BRCA2)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   4.5 kDa
 
UniProt   P51587 (1517-1551)
Sequence   FASTA