High Conformational Flexibility of the E2F1/DP1/DNA Complex.

Saad D, Paissoni C, Chaves-Sanjuan A, Nardini M, Mantovani R, Gnesutta N, Camilloni C, J Mol Biol 433(18):167119 (2021) Europe PMC

SASDKH2 – E2F1/DP1 in complex with a 15-mer DNA oligo

Transcription factor E2F1
15-mer DNA oligo of the human E2F1 promoter
E2F dimerization partner 1
MWexperimental 64 kDa
MWexpected 58 kDa
VPorod 99 nm3
log I(s) 1.08×10-2 1.08×10-3 1.08×10-4 1.08×10-5
Transcription factor E2F1 15-mer DNA oligo of the human E2F1 promoter E2F dimerization partner 1 small angle scattering data  s, nm-1
ln I(s)
Transcription factor E2F1 15-mer DNA oligo of the human E2F1 promoter E2F dimerization partner 1 Guinier plot ln 1.08×10-2 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
Transcription factor E2F1 15-mer DNA oligo of the human E2F1 promoter E2F dimerization partner 1 Kratky plot 1.104 0 3 sRg
p(r)
Transcription factor E2F1 15-mer DNA oligo of the human E2F1 promoter E2F dimerization partner 1 pair distance distribution function Rg: 3.9 nm 0 Dmax: 11.8 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Transcription factor E2F1 15-mer DNA oligo of the human E2F1 promoter E2F dimerization partner 1 CUSTOM IN-HOUSE model

Synchrotron SAXS data from solutions of E2F1/DP1 in complex with a 15-mer DNA oligo in 50 mM Tris-Cl pH 8.0, 150 mM NaCl and 2 mM DTT, pH 8 were collected on the B21 beam line at the Diamond Light Source storage ring (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.0946 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 45.00 μl sample at 4 mg/ml was injected at a 0.08 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 20°C. 20 successive 1 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.

Transcription factor E2F1 (E2F1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   20.5 kDa
 
UniProt   Q01094 (121-301)
Sequence   FASTA
 
15-mer DNA oligo of the human E2F1 promoter
Mol. type   DNA
Olig. state   Monomer
Mon. MW   9.3 kDa
Sequence   FASTA
 
E2F dimerization partner 1 (DP1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   28.7 kDa
 
UniProt   Q14186 (105-353)
Sequence   FASTA