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

SASDQU9 – His-Tagged Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E] in complex with His-tagged fourth BRC repeat (BRC4)

DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged)
Breast cancer type 2 susceptibility protein
MWexperimental 45 kDa
MWexpected 46 kDa
VPorod 78 nm3
log I(s) 4.67×10-2 4.67×10-3 4.67×10-4 4.67×10-5
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein small angle scattering data  s, nm-1
ln I(s)
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein Guinier plot ln 4.68×10-2 Rg: 3.0 nm 0 (3.0 nm)-2 s2
(sRg)2I(s)/I(0)
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein Kratky plot 1.104 0 3 sRg
p(r)
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein pair distance distribution function Rg: 3.4 nm 0 Dmax: 16.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
His-Tagged Monomeric DNA repair protein RAD51 homolog 1 double mutant [F86E, A89E] in complex with His-tagged fourth BRC repeat (BRC4) Rg histogram Rg, nm
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein EOM/RANCH model

log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein ALPHAFOLD model

log I(s)
 s, nm-1
DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) Breast cancer type 2 susceptibility protein PDB (PROTEIN DATA BANK) model

Synchrotron SAXS data from solutions of His-Tagged DNA repair protein RAD51 homolog 1 [F86E, A89E] in complex with the His-tagged fourth BRC repeat of BRC4 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 = 0.095373 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 6.1 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. 36 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 obtained through static light scattering (SLS).

DNA repair protein RAD51 homolog 1 (F86E, A89E, His-Tagged) (HisRAD51 [F86E,A89E])
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   39.8 kDa
 
UniProt   Q06609 (1-339)
Sequence   FASTA
 
Breast cancer type 2 susceptibility protein (HisTaggedBRC4)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   6.3 kDa
 
UniProt   P51587 (1517-1551)
Sequence   FASTA
 
PDB ID   1N0W