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

Harada, Ayaka; Sato, Yukari; Kamimura, Naofumi; Venugopalan, Nagarajan; Masai, Eiji; Senda, Toshiya

doi:10.1107/S2053230X16018665

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

Journal Article · Tue Nov 29 23:00:00 EST 2016 · Acta Crystallographica. Section F, Structural Biology Communications

DOI:https://doi.org/10.1107/S2053230X16018665· OSTI ID:1373907

Harada, Ayaka; Sato, Yukari; Kamimura, Naofumi; Venugopalan, Nagarajan; Masai, Eiji; Senda, Toshiya

A tetrahydrofolate-dependentO-demethylase, LigM, from Sphingobiumsp. SYK-6 was crystallized by the hanging-drop vapour-diffusion method. However, the obtained P3₁21 orP3₂21 crystals, which diffracted to 2.5–3.3 Å resolution, were hemihedrally twinned. To overcome the twinning problem, microseeding using P3₁21/P3₂ 21 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 parametersa= 102.0,b= 117.3,c = 128.1 Å. The P2₁2₁2 crystals diffracted to better than 2.0 Å resolution after optimizing the cryoconditions. Phasing using the single anomalous diffraction method was successful at 3.0 Å resolution with a Pt-derivative crystal. This experience suggested that microseeding is an effective method to overcome the twinning problem, even when twinned crystals are utilized as microseeds.

Research Organization:: Argonne National Laboratory (ANL)

Sponsoring Organization:: Japan Society for the Promotion of Science (JSPS); USDOE Office of Science - Office of Basic Energy Sciences - Scientific User Facilities Division; National Institutes of Health (NIH)

DOE Contract Number:: AC02-06CH11357

OSTI ID:: 1373907

Journal Information:: Acta Crystallographica. Section F, Structural Biology Communications, Journal Name: Acta Crystallographica. Section F, Structural Biology Communications Journal Issue: 12 Vol. 72; ISSN 2053-230X

Publisher:: International Union of Crystallography

Country of Publication:: United States

Language:: English

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tetrahydrofolate

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