Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O-demethylase crystals by the microseeding method.

ABSTRACT: A tetrahydrofolate-dependent O-demethylase, LigM, from Sphingobium sp. SYK-6 was crystallized by the hanging-drop vapour-diffusion method. However, the obtained P3₁21 or P3₂21 crystals, which diffracted to 2.5-3.3 Å resolution, were hemihedrally twinned. To overcome the twinning problem, microseeding using P3₁21/P3₂21 crystals as microseeds was performed with optimization of the reservoir conditions. As a result, another crystal form was obtained. The newly obtained crystal diffracted to 2.5-3.0 Å resolution and belonged to space group P2₁2₁2, with unit-cell parameters a = 102.0, b = 117.3, c = 128.1 Å. The P2₁2₁2 crystals diffracted to better than 2.0 Å resolution after optimizing the cryoconditions. P

SUBMITTER: Harada A 

PROVIDER: S-EPMC5137467 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature