Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum

被引：22
|
作者：
Pasquo, Alessandra [1 ]
Bonamore, Alessandra
Franceschini, Stefano
Macone, Alberto
Boffi, Alberto
Ilari, Andrea
机构：
[1] ENEA, Casaccia Res Ctr, Dipartimento BIOTEC, Sez Genet & Genom Vegetale, I-00100 Rome, Italy
来源：
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2008年 / 64卷
关键词：
D O I：
10.1107/S1744309108005678
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Norcoclaurine synthase ( NCS) catalyzes the condensation of 3,4-dihydroxy-phenylethylamine ( dopamine) and 4-hydroxyphenylacetaldehyde (4-HPAA) as the first committed step in the biosynthesis of benzylisoquinoline alkaloids in plants. The protein was cloned, expressed and purified. Crystals were obtained at 294 K by the hanging-drop vapour-diffusion method using ammonium sulfate and sodium chloride as precipitant agents and diffract to better than 3.0 angstrom resolution using a synchrotron-radiation source. The crystals belong to the trigonal space group P3(1)21, with unit-cell parameters a = b = 86.31, c = 118.36 angstrom. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 angstrom resolution. The model is under construction.
引用
收藏
页码：281 / 283
页数：3
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