Post-translational regulation and trafficking of the granulin-containing protease RD21 of Arabidopsis thaliana.
Post-translational regulation and trafficking of the granulin-containing protease RD21 of Arabidopsis thaliana.
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DOI：
10.1371/journal.pone.0032422
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
van der Hoorn RA
中科院分区：
文献类型：
--
作者：
Gu C;Shabab M;Strasser R;Wolters PJ;Shindo T;Niemer M;Kaschani F;Mach L;van der Hoorn RA
RD21样蛋白酶是普遍存在的植物特异性木瓜蛋白酶样蛋白酶，其典型特征在于携带C-末端颗粒蛋白结构域。RD21样蛋白酶参与免疫，并与衰老和各种类型的生物和非生物胁迫有关。在这里，我们询问拟南芥RD21的调控和贩运的定点诱变，农杆菌渗透，蛋白质印迹，蛋白酶活性分析和蛋白质降解。利用引入的N-聚糖传感器，进行了去糖基化实验和糖工程N。本塞姆植物，我们表明，RD21通过高尔基体，它成为岩藻糖基化。我们的研究表明，RD21在三个翻译后水平受到调控。前结构域去除不阻断在催化Cys突变体，表明RD21被激活的蛋白水解级联。然而，RD21在拟南芥中的激活不需要液泡加工酶（VPE）或糊粉蛋白样蛋白酶AALP。相反，颗粒蛋白结构域的去除需要催化Cys和His残基，因此是自催化的。此外，SDS可以（重新）激活潜伏的RD21在拟南芥叶提取物，表明存在第三层的翻译后调节，可能是由内源性抑制剂介导的。RD21在拟南芥叶提取物中引起显性蛋白酶活性，负责SDS诱导的蛋白质组降解。
RD21-like proteases are ubiquitous, plant-specific papain-like proteases typified by carrying a C-terminal granulin domain. RD21-like proteases are involved in immunity and associated with senescence and various types of biotic and abiotic stresses. Here, we interrogated Arabidopsis RD21 regulation and trafficking by site-directed mutagenesis, agroinfiltration, western blotting, protease activity profiling and protein degradation. Using an introduced N-glycan sensor, deglycosylation experiments and glyco-engineered N. benthamiana plants, we show that RD21 passes through the Golgi where it becomes fucosylated. Our studies demonstrate that RD21 is regulated at three post-translational levels. Prodomain removal is not blocked in the catalytic Cys mutant, indicating that RD21 is activated by a proteolytic cascade. However, RD21 activation in Arabidopsis does not require vacuolar processing enzymes (VPEs) or aleurain-like protease AALP. In contrast, granulin domain removal requires the catalytic Cys and His residues and is therefore autocatalytic. Furthermore, SDS can (re-)activate latent RD21 in Arabidopsis leaf extracts, indicating the existence of a third layer of post-translational regulation, possibly mediated by endogenous inhibitors. RD21 causes a dominant protease activity in Arabidopsis leaf extracts, responsible for SDS-induced proteome degradation.
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影响因子：
4
作者：
Bateman, Andrew;Bennett, Hugh P. J.
通讯作者：
Bennett, Hugh P. J.
影响因子：
27.5
作者：
Schoberer J;Strasser R
通讯作者：
Strasser R
DOI：
10.1007/bf00331014
发表时间：
1986-01-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
KONCZ, C;SCHELL, J
通讯作者：
SCHELL, J
影响因子：
7.2
作者：
Kaschani, Farnusch;Verhelst, Steven H. L.;van der Hoorn, Renier A. L.
通讯作者：
van der Hoorn, Renier A. L.
影响因子：
--
作者：
Greenbaum, D;Medzihradszky, KF;Bogyo, M
通讯作者：
Bogyo, M