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
SASDU33 – Phosphorylated heterochromatin protein HP1α, S97A mutant, full-length (pHP1α_S97A) in 50 mM NaCl
Synchrotron SAXS data from solutions of phosphorylated heterochromatin protein HP1α (S97A mutant, full-length) in 50 mM NaCl in 20 mM sodium phosphate, 50 mM NaCl, 1 mM 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 200.00 μl sample at 14 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. 328 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 48.3 kDa.
s, nm-1
