Structural mechanisms of autoinhibition and substrate recognition by the ubiquitin ligase HACE1

Düring J, Wolter M, Toplak J, Torres C, Dybkov O, Fokkens T, Bohnsack K, Urlaub H, Steinchen W, Dienemann C, Lorenz S, Nature Structural & Molecular Biology (2024) DOI

SASDTC5 – Full-length E3 ubiquitin-protein ligase HACE1

E3 ubiquitin-protein ligase HACE1
MWexperimental 225 kDa
MWexpected 205 kDa
VPorod 379 nm3
log I(s) 2.15×103 2.15×102 2.15×101 2.15×100
E3 ubiquitin-protein ligase HACE1 small angle scattering data  s, nm-1
ln I(s)
E3 ubiquitin-protein ligase HACE1 Guinier plot ln 2.15×103 Rg: 5.2 nm 0 (5.2 nm)-2 s2
(sRg)2I(s)/I(0)
E3 ubiquitin-protein ligase HACE1 Kratky plot 1.104 0 √3 sRg
p(r)
E3 ubiquitin-protein ligase HACE1 pair distance distribution function Rg: 5.1 nm 0 Dmax: 16.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
E3 ubiquitin-protein ligase HACE1 GASBOR model
log I(s)
 s, nm-1
E3 ubiquitin-protein ligase HACE1 ALLOSMOD model
Synchrotron SAXS data from solutions of full-length HACE1 in 50 mM HEPES, 50 mM NaCl, 5 mM DTT, pH 8 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 = 0.123974 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 40.00 μl sample at 9.7 mg/ml was injected onto a Cytiva Superdex 200 Increase 10/300 column at 20°C. 13 successive 0.995 second frames were collected through the SEC elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Flow rate = UNKNOWN

E3 ubiquitin-protein ligase HACE1
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   102.4 kDa
 
UniProt   Q8IYU2 (1-909)
Sequence   FASTA