Crystallization and X-ray diffraction analysis of ornithine cyclodeaminase from Pseudomonas putida

被引:6
作者
Alam, S
Wang, SC
Ruzicka, FJ
Frey, PA
Wedekind, JE [1 ]
机构
[1] Univ Rochester, Sch Med & Dent, Dept Biochem & Biophys, Rochester, NY 14642 USA
[2] Univ Wisconsin, Dept Biochem, Madison, WI 53706 USA
来源
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2004年 / 60卷
关键词
D O I
10.1107/S0907444904005256
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Ornithine cyclodeaminase (OCD) is a member of the mu-crystallin protein family, the biological activity of which is the conversion of L-ornithine to L-proline and ammonia. In order to elucidate the functional groups of this enzyme that are involved in catalysis, the crystallization of OCD from Pseudomonas putida was undertaken. Using microbatch-under-oil screening at the high-throughput crystallization laboratory (HTC) at the Hauptman-Woodward Medical Research Institute Inc. (HWI Buffalo, NY, USA), numerous crystallization conditions were rapidly identified. Several conditions could be reproduced on a larger scale as vapor-diffusion experiments in-house. The best diffraction-quality crystals were obtained from solutions of 40%(v/v) 2-methyl-2,4-pentanediol buffered at pH 6.0 with 0.1 M MES and diffracted X-rays to 1.68 Angstrom resolution. Crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 70.0, b = 78.3, c = 119.4 Angstrom. The V-M was 2.1 Angstrom(3) Da(-1), corresponding to 42% solvent, which is consistent with two 38.5 kDa molecules per asymmetric unit. The structure determination is under way using experimental phasing methods.
引用
收藏
页码:941 / 944
页数:4
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