Open AccessCloning, expression, crystallization and preliminary X‐ray data analysis of norcoclaurine synthase from Thalictrum flavum
Author(s) -
Pasquo Alessandra,
Bonamore Alessandra,
Franceschini Stefano,
Macone Alberto,
Boffi Alberto,
Ilari Andrea
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108005678
Subject(s) - benzylisoquinoline , crystallization , biosynthesis , chemistry , atp synthase , crystallography , cloning (programming) , resolution (logic) , ammonium sulfate , stereochemistry , derivative (finance) , biochemistry , enzyme , organic chemistry , artificial intelligence , computer science , programming language , financial economics , economics
Norcoclaurine synthase (NCS) catalyzes the condensation of 3,4‐dihydroxyphenylethylamine (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 Å resolution using a synchrotron‐radiation source. The crystals belong to the trigonal space group P 3 1 21, with unit‐cell parameters a = b = 86.31, c = 118.36 Å. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 Å resolution. The model is under construction.