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SorvallTM RC 6 Plus Centrifuge

Sorvall TM RC 6 Plus离心机

Company: Thermo Fisher Scientific
Catalog#: RC 6 Plus
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Heterologous Expression and Purification of Catalytic Domain of CESA1 from Arabidopsis thaliana
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2016-10-20
[Abstract]  Heterologous expression of plant cellulose synthase (CESA) and its purification has remained a challenge for decades impeding detailed biophysical, biochemical and structural characterization of this key enzyme. An in-depth knowledge of structure and function of CESA proteins would enable us to better understand the hierarchical structure of the plant cell wall. Here, we report a detailed, and reproducible method of purification of catalytic domain of CESA1 from Arabidopsis thaliana that was recombinantly expressed in Escherichia coli. The method relies on a two stage purification procedure to obtain the catalytic domain in monomer and trimer forms. The biochemical and biophysical data including low resolution structures of the protein have been published (Vandavasi et ... [摘要]  植物纤维素合酶(CESA)的异源表达及其纯化在数十年来一直是阻碍该关键酶的详细生物物理,生物化学和结构表征的挑战。对CESA蛋白的结构和功能的深入了解将使我们能够更好地理解植物细胞壁的层次结构。在这里,我们报告了在大肠杆菌中重组表达的来自拟南芥的CESA1的催化结构域的详细的和可重复的纯化方法。该方法依赖于两阶段纯化方法以获得单体和三聚体形式的催化结构域。已经公开了包括蛋白质的低分辨率结构的生物化学和生物物理数据(Vandavasi等人,2016)。目前,这种蛋白质的结晶研究正在进行中。

[Backg 圆形] 纤维素是植物细胞壁最重要的结构组分,地球最大的生物可再生材料来源,但它的植物合成机制了解很少。植物纤维素合成复合物(CSC),也称为"玫瑰花结",因为其在电子显微镜图像中的六聚体外观,是一个大的多亚基跨膜蛋白复合物负责纤维素链的合成和它们组装成微原纤维。 ...

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