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西葫芦髓部中马维花青素的结晶及初步X射线晶体学研究。

Crystallization and preliminary X-ray crystallographic studies of mavicyanin from Cucurbita pepo medullosa.

作者信息

Xie Yong, Inoue Tsuyoshi, Miyamoto Youichi, Matsumura Hiroyoshi, Kataoka Kunishige, Yamaguchi Kazuya, Suzuki Shinnichiro, Kai Yasushi

机构信息

Department of Materials Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.

出版信息

Acta Crystallogr D Biol Crystallogr. 2003 Aug;59(Pt 8):1474-6. doi: 10.1107/s0907444903011326. Epub 2003 Jul 23.

Abstract

Mavicyanin isolated from Cucurbita pepo medullosa is a glycosylated protein containing a single polypeptide chain of 109 amino-acid residues and is a member of the phytocyanin subclass of cupredoxins. Non-glycosylated recombinant mavicyanin, which was expressed in Escherichia coli, was crystallized by the hanging-drop vapour-diffusion method with ammonium sulfate as the precipitant at pH 5.5. The crystals belonged to the hexagonal space group P6(1) (or P6(5)), with unit-cell parameters a = 64.0, c = 245.0 A, four molecules per asymmetric unit and a solvent content of 59%. X-ray diffraction data were collected to 1.6 A resolution. To solve the structure of mavicyanin, the MAD method as well as a Patterson search method using the structure of stellacyanin as a starting model are presently being utilized.

摘要

从西葫芦髓质中分离出的马维花青素是一种糖基化蛋白,含有一条由109个氨基酸残基组成的单多肽链,属于铜蓝蛋白中植物花青素亚类的成员。在大肠杆菌中表达的非糖基化重组马维花青素,通过悬滴气相扩散法,以硫酸铵作为沉淀剂,在pH 5.5条件下结晶。晶体属于六方空间群P6(1)(或P6(5)),晶胞参数a = 64.0,c = 245.0 Å,每个不对称单元有四个分子,溶剂含量为59%。X射线衍射数据收集到1.6 Å分辨率。为了解决马维花青素的结构,目前正在使用MAD方法以及以星蓝蛋白结构为起始模型的帕特森搜索方法。

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