Furukawa A,
Yonezawa K,
Negami T,
Yoshimura Y,
Hayashi A,
Nakayama J,
Adachi N,
Senda T,
Shimizu K,
Terada T,
Shimizu N,
Nishimura Y,
Nucleic Acids Research
53(6)
(2025)
DOI
SASDU53 – Phosphorylated heterochromatin protein HP1α, S97A mutant, full-length (pHP1α_S97A) in 500 mM NaCl
Synchrotron SAXS data from solutions of phosphorylated heterochromatin protein HP1α (S97A mutant, full-length) in 500 mM NaCl in 20 mM sodium phosphate, 500 mM NaCl, 1mM DTT, pH 7 were collected on the BL-10C beam line at the Photon Factory (PF), High Energy Accelerator Research Organization (KEK; Tsukuba, Japan) using a Pilatus3 2M detector at a sample-detector distance of 2.0 m and at a wavelength of λ = 0.1213 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 250.00 μl sample at 5 mg/ml was injected onto a Cytiva Superdex 200 Increase 10/300 column at 20°C. The flow rate was set to 0.05 mL/min around the peak region. 466 successive 20 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.
SEC-SAXS experiment was performed using an HPLC system, Prominence-i (SHIMADZU). SEC-MALS experiments were performed before the SEC-SAXS experiments and the molecular weight was estimated to be 44.5 kDa.
s, nm-1
