Unorthodox PCNA Binding by Chromatin Assembly Factor 1

Gopinathan Nair A, Rabas N, Lejon S, Homiski C, Osborne M, Cyr N, Sverzhinsky A, Melendy T, Pascal J, Laue E, Borden K, Omichinski J, Verreault A, International Journal of Molecular Sciences 23(19):11099 (2022) DOI

SASDP79 – Chromatin assembly factor 1 - long alpha-helix domain

Chromatin assembly factor 1 subunit A
MWexperimental 35 kDa
MWexpected 18 kDa
VPorod 54 nm3
log I(s) 3.85×10-3 3.85×10-4 3.85×10-5 3.85×10-6
Chromatin assembly factor 1 subunit A small angle scattering data  s, nm-1
ln I(s)
Chromatin assembly factor 1 subunit A Guinier plot ln 3.86×10-3 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Chromatin assembly factor 1 subunit A Kratky plot 1.104 0 √3 sRg
p(r)
Chromatin assembly factor 1 subunit A pair distance distribution function Rg: 4.6 nm 0 Dmax: 17.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Chromatin assembly factor 1 subunit A DAMMIN model
SAXS data from solutions of the chromatin assembly factor 1 - long alpha-helix domain in 20 mM sodium phosphate, 200 mM NaCl, 0.1 mM TCEP, pH 7 were collected using a Xenocs BioXolver L with MetalJet instrument (Département de Biochimie, Université de Montréal, Canada) equipped with a Pilatus3 R 300K detector at a sample-detector distance of 600 m and at a wavelength of λ = 0.134 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 500.00 μl sample at 5.8 mg/ml was injected at a 0.05 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 160 successive 60 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.

Chromatin assembly factor 1 subunit A (p150L)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   17.6 kDa
 
UniProt   Q13111 (342-475)
Sequence   FASTA