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

SASDTD5 – N-terminally truncated E3 ubiquitin-protein ligase HACE1

E3 ubiquitin-protein ligase HACE1
MWexperimental 118 kDa
MWexpected 100 kDa
VPorod 219 nm3
log I(s) 1.92×103 1.92×102 1.92×101 1.92×100
E3 ubiquitin-protein ligase HACE1 small angle scattering data  s, nm-1
ln I(s)
E3 ubiquitin-protein ligase HACE1 Guinier plot ln 1.92×103 Rg: 4.6 nm 0 (4.6 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: 4.7 nm 0 Dmax: 17.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
E3 ubiquitin-protein ligase HACE1 MULTIFOXS model
E3 ubiquitin-protein ligase HACE1 MULTIFOXS model
E3 ubiquitin-protein ligase HACE1 MULTIFOXS model
Synchrotron SAXS data from solutions of N-terminally truncated 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. 17 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   Monomer
Mon. MW   99.8 kDa
 
UniProt   Q8IYU2 (22-909)
Sequence   FASTA