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