Activity-based protein profiling of infected plants.
Activity-based protein profiling of infected plants.
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受感染植物的基于活性的蛋白质分析。
DOI：
10.1007/978-1-61779-501-5_3
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发表时间：
2012
影响因子：
--
通讯作者：
R. V. D. van der Hoorn
中科院分区：
文献类型：
--
作者：
F. Kaschani;C. Gu;R. V. D. van der Hoorn
基于活性的蛋白质谱（ABPP）是一种检测和比较蛋白质组中蛋白质活性的有效分析方法。这是使用基于特异性活性的探针来实现的，所述探针通常衍生自抑制剂并且分别连接至如罗丹明或生物素的报告基团以用于荧光检测和/或亲和纯化。探针与蛋白质的活性位点残基反应并共价且不可逆地连接，从而促进标记蛋白质的分离、检测和鉴定。在本协议中，我们描述了标记，纯化和鉴定标记的蛋白质从凝胶所需的所有步骤，并显示如何在两个蛋白质组的活动可以比较。使用三功能探针TriFP鉴定来自感染灰葡萄孢的拟南芥植物的丝氨酸水解酶作为实例。
Activity-based protein profiling (ABPP) is a powerful analytical method to detect and compare the activity of proteins in proteomes. This is achieved using specific activity-based probes that are often derived from inhibitors and are linked to reporter groups like rhodamine or biotin for fluorescence detection and/or affinity purification, respectively. The probes react with the active site residue of proteins and become covalently and irreversibly attached, facilitating the separation, detection and identification of the labelled proteins. In this protocol we describe all the steps required for labelling, purification and identification of labelled proteins from gels and show how activities in two proteomes can be compared. The identification of serine hydrolases from Arabidopsis plants infected with Botrytis cinerea using the trifunctional probe TriFP is used as an example.
在蛋白质微序列分析之前对玻璃纤维上的胱氨酸残基进行原位烷基化的程序。
影响因子：
2.9
作者：
Andrews,PC;Dixon,JE
通讯作者：
Dixon,JE