Multidomain architecture of estrogen receptor reveals interfacial cross-talk between its DNA-binding and ligand-binding domains.

Huang W, Peng Y, Kiselar J, Zhao X, Albaqami A, Mendez D, Chen Y, Chakravarthy S, Gupta S, Ralston C, Kao HY, Chance MR, Yang S, Nat Commun 9(1):3520 (2018) PubMed

SASDDU8 – Multidomain architecture of the estrogen receptor reveals interfacial cross-talk between its DNA-binding and ligand-binding domains

Estrogen receptor
ERE1
ERE2
Estradiol
hERa peptide1
hERa peptide2
MWexperimental 98 kDa
MWexpected 100 kDa
log I(s) 4.10×100 4.10×10-1 4.10×10-2 4.10×10-3
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 small angle scattering data  s, nm-1
ln I(s)
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 Guinier plot ln 4.10×100 Rg: 3.8 nm 0 (3.8 nm)-2 s2
(sRg)2I(s)/I(0)
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 Kratky plot 1.104 0 √3 sRg
p(r)
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 pair distance distribution function Rg: 3.8 nm 0 Dmax: 11.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 CUSTOM IN-HOUSE model
log I(s)
 s, nm-1
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 CUSTOM IN-HOUSE model
log I(s)
 s, nm-1
Estrogen receptor ERE1 ERE2 Estradiol hERa peptide1 hERa peptide2 CUSTOM IN-HOUSE model
Synchrotron SAXS data from solutions of the multi-component estrogen receptor in 10 mM CHES, 125 mM NaCl, 5mM KCl, 4 mM MgCl2, 50 mM arginine, 50 mM glutamate, 5 mM TCEP, 5% glycerol, 10 µm Zn acetate, 10 µM estradiol, pH 9.5 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS; Argonne, IL, USA) using a MAR 165 CCD detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.103 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.1 and 0.8 mg/ml were measured at 10°C from size-exclusion chromatography eluates. 20 successive 1.100 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.

Additional SEC parameters: Column type: GE Healthcare Superdex 200 GL 10/300; Flow rate: 0.5 ml/min; Sample injection concentration: 3.5 mg/ml; Injection volume: 500 µl.

Estrogen receptor (hERa CDE)
Mol. type   Protein
Olig. state   Dimer
Mon. MW   42.5 kDa
 
UniProt   P03372 (181-552)
Sequence   FASTA
 
ERE1 (hERa_ERE1)
Mol. type   DNA
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   5.9 kDa
Sequence   FASTA
 
ERE2 (hERa_ERE2)
Mol. type   DNA
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   5.9 kDa
Sequence   FASTA
 
Estradiol
Mol. type   Other
Mon. MW   0.3 kDa
Sequence   FASTA
 
hERa peptide1
Mol. type   Protein
Olig. state   Monomer
Mon. MW   1.7 kDa
Sequence   FASTA
 
hERa peptide2
Mol. type   Protein
Olig. state   Monomer
Mon. MW   1.7 kDa
Sequence   FASTA