Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase

Lu Heshu; Yang Mingfeng; Liu Chunmei; Lu Ping; Cang Huaixing; Ma Lanqing

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Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase

机译：虎杖双功能查尔酮合酶/苯扎丙酮合酶的蛋白制备，结晶及X射线初步分析

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The chalcone synthase (CHS) superfamily of type III polyketide synthases (PKSs) generate the backbones of a variety of plant secondary metabolites. An active bifunctional chalcone synthase/benzalacetone synthase (CHS/BAS) from Polygonum cuspidatum was overexpressed in Escherichia coli as a C-terminally polyhistidine-tagged fusion protein, purified to homogeneity and crystallized using polyethylene glycol 4000 as a precipitant. The production of well shaped crystals of the complex between PcPKS1 and benzalacetone was dependent on the presence of sorbitol and barium chloride as additives. The crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 80.23, b = 81.01, c = 122.89 angstrom, and diffracted X-rays to at least 2.0 angstrom resolution.

机译：III型聚酮化合物合酶（PKS）的查尔酮合酶（CHS）超家族可产生多种植物次生代谢产物的骨架。来自虎杖的一种活性双功能查尔酮合酶/苯并丙酮合酶（CHS / BAS）在大肠杆菌中以C端多聚组氨酸标记的融合蛋白过表达，纯化至均质并使用聚乙二醇4000作为沉淀剂结晶。 PcPKS1与苯并丙酮之间的复合物形状良好的晶体的生产取决于山梨糖醇和氯化钡作为添加剂的存在。晶体属于正交晶体空间群P2（1）2（1）2（1），单位晶胞参数a = 80.23，b = 81.01，c = 122.89埃，并且将X射线衍射到至少2.0埃的分辨率。

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《Acta crystallographica, Section F. Structural biology and crystallization communications》 |2013年第8期|共5页
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Lu Heshu; Yang Mingfeng; Liu Chunmei; Lu Ping; Cang Huaixing; Ma Lanqing;
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中图分类晶体学;
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1. Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase [J] . Lu Heshu, Yang Mingfeng, Liu Chunmei, Acta crystallographica, Section F. Structural biology and crystallization communications . 2013,第8期

机译：虎杖双功能查尔酮合酶/苯扎丙酮合酶的蛋白制备，结晶及X射线初步分析
2. Preparation, Crystallization and Preliminary X-Ray Analysis of Threonine Synthase from Streptococcus mutans [J] . De-Wei Tang, Lan-Fen Li, Ya-Mei Yu, Protein and peptide letters . 2007,第8期

机译：变形链球菌苏氨酸合酶的制备，结晶及X射线初步分析
3. Heterologous expression, purification, crystallization and preliminary X-ray diffraction analysis of KDO8P synthase from Arabidopsis thaliana [J] . Fengxue Zhang, Yingwu Xu, Zhijun Zhang Protein Expression and Purification . 2014,第Null期

机译：拟南芥KDO8P合酶的异源表达，纯化，结晶和初步X射线衍射分析
4. Molecular Cloning and Sequences Analysis of Chalcone Synthase Gene from Fagopyrum Tataricum [C] . Huangyuan She, Shaohong He, Zhi Zhou, International conference on informatics and management science;IMS 2012 . 2013

机译：苦荞麦查尔酮合酶基因的分子克隆与序列分析
5. An investigation into plant type III polyketide synthases: A styrylpyrone synthase from Equisetum hyemale and anther-specific chalcone synthase-like enzymes from Physcomitrella patens and Arabidopsis thaliana. [D] . Colpitts, Che Caswell. 2009

机译：植物III型聚酮化合物合酶的研究：马齿Equi的苯乙烯基吡喃酮合酶和帕氏假单胞菌和拟南芥的花药特异性查尔酮合酶样酶。
6. Protein preparation crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase [O] . Heshu Lu, Mingfeng Yang, Chunmei Liu, 2013

机译：虎杖双功能查尔酮合酶/苯扎丙酮合酶的蛋白质制备结晶及初步X射线分析
7. Characterization, crystallization and preliminary X-ray analysis of bifunctional dihydrofolate reductase–thymidylate synthase fromPlasmodium falciparum [O] . Penchit Chitnumsub, Jirundon Yavaniyama, Jarunee Vanichtanankul, 2004

机译：双官能二氢酚酸盐还原酶 - 胸苷合酶的表征，结晶及初步X射线分析从vlimodium falciparum

Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase

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