Mechanistic insights into enhancement or inhibition of phase separation by different polyubiquitin chains.

Dao TP, Yang Y, Presti MF, Cosgrove MS, Hopkins JB, Ma W, Loh SN, Castañeda CA, EMBO Rep :e55056 (2022) Europe PMC

SASDPA3 – K63-linked tetraubiquitin (Ub4)

Polyubiquitin-B
MWexperimental 35 kDa
MWexpected 34 kDa
VPorod 44 nm3
log I(s) 3.26×10-2 3.26×10-3 3.26×10-4 3.26×10-5
Polyubiquitin-B small angle scattering data  s, nm-1
ln I(s)
Polyubiquitin-B Guinier plot ln 3.26×10-2 Rg: 3.2 nm 0 (3.2 nm)-2 s2
(sRg)2I(s)/I(0)
Polyubiquitin-B Kratky plot 1.104 0 3 sRg
Dmax: 13 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of K63-linked tetraubiquitin (Ub4) in 20 mM sodium phosphate, 0.5 mM EDTA, 0.02% NaN3, pH 6.8 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.1033 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 300.00 μl sample at 7 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 2000 successive 0.500 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.

K63-linked tetraubiquitin with distal Ub as K63R Ub; all other units are Ub. Ub units are connected by K63-linkages. Chain was made enzymatically.

Polyubiquitin-B (K63-Ub4)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   34.1 kDa
 
UniProt   P0CG47 (1-76)
Sequence   FASTA