Polyubiquitin ligand-induced phase transitions are optimized by spacing between ubiquitin units

Galagedera S Dao T, Enos S, Chaudhuri A, Schmit J, CastaƱeda C, Proceedings of the National Academy of Sciences 120(42) (2023) DOI

SASDRD7 – HOTag6 tetramerization domain followed by a Gly-Ser linker fused to human monoubiquitin with 200 mM NaCl

HOTag6-(GS)-Ubiquitin
MWI(0) 49 kDa
MWexpected 51 kDa
VPorod 70 nm3
log I(s) 1.10×10-2 1.10×10-3 1.10×10-4 1.10×10-5
HOTag6-(GS)-Ubiquitin small angle scattering data  s, nm-1
ln I(s)
HOTag6-(GS)-Ubiquitin Guinier plot ln 1.10×10-2 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
HOTag6-(GS)-Ubiquitin Kratky plot 1.104 0 3 sRg
p(r)
HOTag6-(GS)-Ubiquitin pair distance distribution function Rg: 3.8 nm 0 Dmax: 12.6 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of HOTag6 tetramerization domain followed by a Gly-Ser linker fused to human monoubiquitin with 200 mM NaCl in 20 mM sodium phosphate, 200 mM NaCl, 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 storage ring (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). One solute concentration of 4.68 mg/ml was measured at 20°C. 58 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.

HOTag6 is a ~30-amino-acid tetramerization domain followed by a Gly-Ser linker followed by the sequence for human monoubiquitin (UniProt ID: P0CG47, https://www.uniprot.org/uniprot/P0CG47, amino acids 1-76) . This is a designed protein (no natural variant exists). The HOTag6 sequence is from the Shu group at the University of California, San Francisco (https://doi.org/10.1016/j.molcel.2017.12.008).

HOTag6-(GS)-Ubiquitin
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   12.8 kDa
Sequence   FASTA