Decoding optimal ligand design for multicomponent condensates

Sarasi Galagedera.

SASDRA7 – HOTag6 tetramerization domain followed by a (Pro-Ala)2 linker fused to human monoubiquitin

HOTag6-(PA)2-Ubiquitin
MWI(0) 50 kDa
MWexpected 52 kDa
VPorod 74 nm3
log I(s) 1.55×10-2 1.55×10-3 1.55×10-4 1.55×10-5
HOTag6-(PA)2-Ubiquitin small angle scattering data  s, nm-1
ln I(s)
HOTag6-(PA)2-Ubiquitin Guinier plot ln 1.56×10-2 Rg: 3.8 nm 0 (3.8 nm)-2 s2
(sRg)2I(s)/I(0)
HOTag6-(PA)2-Ubiquitin Kratky plot 1.104 0 √3 sRg
p(r)
HOTag6-(PA)2-Ubiquitin pair distance distribution function Rg: 4.0 nm 0 Dmax: 15 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of HOTag6 tetramerization domain followed by a (Pro-Ala)2 linker fused to human monoubiquitin 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 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 4.61 mg/ml was measured at 20°C. Five 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 (Pro-Ala)2 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-(PA)2-Ubiquitin
Mol. type   Protein
Organism   synthetic construct
Olig. state   Tetramer
Mon. MW   13.0 kDa
Sequence   FASTA