Diversity of Serine Hydrolase Activities of Unchallenged and Botrytis-infected Arabidopsis thaliana
Diversity of Serine Hydrolase Activities of Unchallenged and Botrytis-infected Arabidopsis thaliana
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DOI：
10.1074/mcp.m800494-mcp200
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发表时间：
2009-05-01
影响因子：
7
通讯作者：
van der Hoorn, Renier A. L.
中科院分区：
文献类型：
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作者：
Kaschani, Farnusch;Gu, Christian;van der Hoorn, Renier A. L.
基于活性的蛋白质图谱是一种显示蛋白质组酶活性的有效方法，它提供了关于酶活性的关键信息，而不是蛋白质或转录本的丰度。我们应用基于活性的蛋白质谱，使用基于氟磷酸盐的探针来显示模式植物拟南芥中丝氨酸水解酶的活性。多维蛋白质鉴定技术和凝胶内分析氟磷标记的叶提取物发现了50多种丝氨酸水解酶，包括数十种蛋白酶、酯酶和脂肪酶，代表10多个不同的酶家族。除了一些功能较好的丝氨酸水解酶，如枯草杆菌枯草杆菌酶TPP2和ARA12、脯氨酸寡肽酶酰基氨基酸释放酶、丝氨酸羧肽酶SNG1和BRS1、羧酸酯酶CXE12、甲基酯酶MES2和MES3以及S-甲酰谷胱甘肽水解酶外，大多数丝氨酸水解酶以前都没有被描述过。我们对瞬时表达的SNG1进行了研究，并对感染了真菌病原菌灰霉病菌的植株进行了调查。除了在灰霉病菌侵染过程中几种拟南芥丝氨酸水解酶活性的下调外，我们还检测到了被认为与致病性有关的灰霉病菌衍生的角质酶和脂肪酶的活性。《分子与细胞蛋白质组学》8：1082-1093,2009。
Activity-based protein profiling is a powerful method to display enzyme activities in proteomes and provides crucial information on enzyme activity rather than protein or transcript abundance. We applied activity-based protein profiling using fluorophosphonate-based probes to display the activities of Ser hydrolases in the model plant Arabidopsis thaliana. Multidimensional protein identification technology and in-gel analysis of fluorophosphonate-labeled leaf extracts revealed over 50 Ser hydrolases, including dozens of proteases, esterases, and lipases, representing over 10 different enzyme families. Except for some well characterized Ser hydrolases like subtilases TPP2 and ARA12, prolyl oligopeptidase acylamino acid-releasing enzyme, serine carboxypeptidase-like SNG1 and BRS1, carboxylesterase-like CXE12, methylesterases MES2 and MES3, and S-formylglutathione hydrolase, the majority of these serine hydrolases have not been described before. We studied transiently expressed SNG1 and investigated plants infected with the fungal pathogen Botrytis cinerea. Besides the down-regulation of several Arabidopsis Ser hydrolase activities during Botrytis infection, we detected the activities of Botrytis-derived cutinases and lipases, which are thought to contribute to pathogenicity. Molecular & Cellular Proteomics 8: 1082-1093, 2009.