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

Abstract

A tetrahydrofolate-dependentO-demethylase, LigM, from Sphingobiumsp. SYK-6 was crystallized by the hanging-drop vapour-diffusion method. However, the obtained P3 121 orP3 221 crystals, which diffracted to 2.5–3.3 Å resolution, were hemihedrally twinned. To overcome the twinning problem, microseeding using P3 121/P3 2 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 P2 12 12, with unit-cell parametersa= 102.0,b= 117.3,c = 128.1 Å. The P2 12 12 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.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
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)
OSTI Identifier:
1373907
DOE Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F, Structural Biology Communications; Journal Volume: 72; Journal Issue: 12
Country of Publication:
United States
Language:
English
Subject:
O-demethylase; Sphingobium sp. SYK-6; cryo-protectant; hemihedral twinning; micro-seeding; tetrahydrofolate

Citation Formats

Harada, Ayaka, Sato, Yukari, Kamimura, Naofumi, Venugopalan, Nagarajan, Masai, Eiji, and Senda, Toshiya. Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O -demethylase crystals by the microseeding method. United States: N. p., 2016. Web. doi:10.1107/S2053230X16018665.
Harada, Ayaka, Sato, Yukari, Kamimura, Naofumi, Venugopalan, Nagarajan, Masai, Eiji, & Senda, Toshiya. Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O -demethylase crystals by the microseeding method. United States. doi:10.1107/S2053230X16018665.
Harada, Ayaka, Sato, Yukari, Kamimura, Naofumi, Venugopalan, Nagarajan, Masai, Eiji, and Senda, Toshiya. Wed . "Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O -demethylase crystals by the microseeding method". United States. doi:10.1107/S2053230X16018665.
@article{osti_1373907,
title = {Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O -demethylase crystals by the microseeding method},
author = {Harada, Ayaka and Sato, Yukari and Kamimura, Naofumi and Venugopalan, Nagarajan and Masai, Eiji and Senda, Toshiya},
abstractNote = {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.},
doi = {10.1107/S2053230X16018665},
journal = {Acta Crystallographica. Section F, Structural Biology Communications},
number = 12,
volume = 72,
place = {United States},
year = {Wed Nov 30 00:00:00 EST 2016},
month = {Wed Nov 30 00:00:00 EST 2016}
}