Abstract
The HsdR subunit of the type I restriction-modification system EcoR124I is responsible for the translocation as well as the restriction activity of the whole complex consisting of the HsdR, HsdM and HsdS subunits, and while crystal structures are available for the wild type and several mutants, the C-terminal domain comprising approximately 150 residues was not resolved in any of these structures. Here, three fusion constructs with the GFP variant pHluorin developed to overexpress, purify and crystallize the C-terminal domain of HsdR are reported. The shortest of the three encompassed HsdR residues 887-1038 and yielded crystals that belonged to the orthorhombic space group C2221, with unit-cell parameters a = 83.42, b = 176.58, c = 126.03 Å, α = β = γ = 90.00° and two molecules in the asymmetric unit (V M = 2.55 Å3 Da-1, solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45 Å.
| Original language | English |
|---|---|
| Journal | Acta Crystallographica Section:F Structural Biology Communications |
| Volume | 72 |
| Pages (from-to) | 672-676 |
| Number of pages | 5 |
| DOIs | |
| Publication status | Published - 01.09.2016 |
Funding
The authors gratefully acknowledge support from the Czech Science Foundation (P207/12/2323; http://www.gacr.cz/); NL was supported by the REU Site Molecular Biophysics award (REU1358737) to Jannette Carey, Princeton University, USA. Access to instruments and other facilities was supported by the Czech Research Infrastructure for Systems Biology C4SYS (project No. LM2015055). Data collection was carried out at the PETRA III light source at DESY, a member of the Helmholtz Association (HGF). The authors would like to thank Dr Johanna Kallio for assistance in using beamline P13.
