Structural basis of dimerization and nucleic acid binding of human DBHS proteins NONO and PSPC1.

Knott GJ, Chong YS, Passon DM, Liang XH, Deplazes E, Conte MR, Marshall AC, Lee M, Fox AH, Bond CS, Nucleic Acids Res (2021) Europe PMC

SASDMT6 – Non-POU domain-containing octamer-binding protein (NONO homodimer) bound to an antisense oligonucleotide

Non-POU domain-containing octamer-binding protein
5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer
MWexperimental 86 kDa
MWexpected 89 kDa
VPorod 153 nm3
log I(s) 6.64×10-2 6.64×10-3 6.64×10-4 6.64×10-5
Non-POU domain-containing octamer-binding protein 5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer small angle scattering data  s, nm-1
ln I(s)
Non-POU domain-containing octamer-binding protein 5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer Guinier plot ln 6.64×10-2 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Non-POU domain-containing octamer-binding protein 5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer Kratky plot 1.104 0 √3 sRg
p(r)
Non-POU domain-containing octamer-binding protein 5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer pair distance distribution function Rg: 4.2 nm 0 Dmax: 18.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Non-POU domain-containing octamer-binding protein 5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer SASREF model
Synchrotron SAXS data from solutions of the NONO homodimer bound to an antisense oligonucleotide in 20 mM Tris-Cl (pH 7.5), 250 mM KCl, 50 mM L-proline, 0.5 mM EDTA were collected on the SAXS/WAXS beam line at the Australian Synchrotron (Melbourne, Australia) using a Pilatus 1M detector at a sample-detector distance of 3.5 m and at a wavelength of λ = 0.1 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 200.00 μl sample at 14 mg/ml was injected at a 0.50 ml/min flow rate onto a Wyatt WTC-030S5 5μm 300+, 7.8x300 mm column at 4°C. 500 successive 1 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.

Non-POU domain-containing octamer-binding protein (NONO)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   30.1 kDa
 
UniProt   Q15233 (53-312)
Sequence   FASTA
 
5-10-5 gapmer phosphorothioate antisense oligonucleotide tetramer (ASO tetramer)
Mol. type   Other
Olig. state   Tetramer
Mon. MW   7.1 kDa
Chemical formula