Molecular basis of the regulation of full-length Nedd4-2 through calcium and 14-3-3 proteins

Veronika Obšilová.

SASDUF3 – Phosphorylated E3 ubiquitin-protein ligase Nedd4-2 complexed to 14-3-3 eta in the presence of calcium

14-3-3 protein eta
Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like
MWexperimental 186 kDa
MWexpected 167 kDa
VPorod 290 nm3
log I(s) 1.50×10-1 1.50×10-2 1.50×10-3 1.50×10-4
14-3-3 protein eta Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like small angle scattering data  s, nm-1
ln I(s)
14-3-3 protein eta Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like Guinier plot ln 1.51×10-1 Rg: 5.5 nm 0 (5.5 nm)-2 s2
(sRg)2I(s)/I(0)
14-3-3 protein eta Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like Kratky plot 1.104 0 3 sRg
p(r)
14-3-3 protein eta Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like pair distance distribution function Rg: 5.7 nm 0 Dmax: 23.5 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of phosphorylated E3 ubiquitin-protein ligase Nedd4-2 complexed to 14-3-3 protein eta in 20 mM HEPES, 150 mM NaCl, 1 mM TCEP, 1 mM CaCl2, 3% glycerol, pH 7.5 were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.123982 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 9.6 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 1800 successive 0.495 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.

14-3-3 protein eta (14-3-3 eta human)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   28.4 kDa
 
UniProt   Q04917 (1-246)
Sequence   FASTA
 
Isoform 5 of E3 ubiquitin-protein ligase NEDD4-like (Nedd4-2 iso5 human)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   110.3 kDa
 
UniProt   Q96PU5-5 (1-955)
Sequence   FASTA