| Detection of Mitogen-activated Protein Kinase (MAPK) Activation upon Exogenous Chemical Application in Arabidopsis Protoplasts
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Author:
Date:
2015-09-20
[Abstract] The mitogen activated protein kinase cascade is a highly conserved signal pathway in plants. The exogenous chemicals, like hormones, can trigger a series of signalling cascades, including MAPK pathway, to modulate the plant physiology. Upon activation, some MAPKs are phosphorylated. It is important to develop methods that can detect changes in the phosphorylation status of MAPKs in plants when they come in contact with external chemicals. This method describes the exogenous treatment of Arabidopsis protoplasts with Kinetin and subsequent detection of the activated MAPKs. This method is useful for studying the effect of exogenously applied chemical compounds on the MAPK signaling cascade in Arabidopsis.
[摘要] 丝裂原活化蛋白激酶级联是植物中高度保守的信号通路。 外源化学物质，如激素，可以触发一系列信号级联，包括MAPK途径，以调节植物生理。 在激活时，一些MAPK被磷酸化。 开发能够检测植物中MAPK在与外部化学品接触时磷酸化状态的变化的方法是重要的。 该方法描述了用激动素对随后检测激活的MAPK的拟南芥原生质体的外源处理。 该方法可用于研究外源施加的化合物对拟南芥中MAPK信号级联的影响。
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| Biotinylation and Purification of Surface-exposed Helicobacter pylori Proteins
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Author:
Date:
2015-04-20
[Abstract] Interactions between pathogenic bacteria and host cells are often mediated by proteins found on the surfaces of the bacteria. The Gram-negative bacterium Helicobacter pylori is predicted to produce at least 50 surface-exposed outer membrane proteins, but there has been relatively little progress in experimentally analyzing the cell-surface proteome of this organism. Herein, we describe in detail a protocol that allows biotinylation and purification of surface-exposed H. pylori proteins. A comparative analysis of surface-exposed proteins identified by this biotinylation-based approach and by several other independent methods is described in a recent publication (Voss et al., 2014).
[摘要] 病原细菌和宿主细胞之间的相互作用通常由在细菌表面上发现的蛋白介导。 革兰氏阴性细菌幽门螺杆菌预计产生至少50个表面暴露的外膜蛋白，但是在实验分析该生物体的细胞表面蛋白质组中相对较少的进展。 在本文中，我们详细描述了允许表面暴露的H的生物素化和纯化的方案。 幽门螺杆菌蛋白。 在最近的出版物（Voss等人，2014）中描述了通过这种基于生物素化的方法和几种其它独立方法鉴定的表面暴露蛋白的比较分析。
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