The fronds tonoplast quantitative proteomic analysis in arsenic hyperaccumulator Pteris vittata L.
The fronds tonoplast quantitative proteomic analysis in arsenic hyperaccumulator Pteris vittata L.
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砷超积累植物 Pteris vittata L 的叶液泡膜定量蛋白质组学分析。
DOI：
10.1016/j.jprot.2014.01.029
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发表时间：
2014-06
影响因子：
3.3
通讯作者：
Ma M
中科院分区：
文献类型：
--
作者：
Shen HL;He ZY;Yan HL;Xing ZN;Chen YS;Xu WX;Xu WZ;Ma M
摘要文章报道了世界上第一种砷超积累植物-蜈蚣蕨（Pteris vittata）的研究进展，发现其地上部分对砷的积累量很高，而根系对砷的积累量很低。以往的研究表明，砷的液泡区室化可能在砷的超积累中起重要作用。vittata，但砷运输到液泡的机制在很大程度上是未知的。我们从叶中分离出液泡膜。采用碘克沙醇分步梯度离心法，在-1mM砷酸盐条件下培养3周，然后采用TMPP蛋白标记技术，结合液相色谱-线性离子阱-轨道阱混合质谱仪分析，对蛋白质进行定量检测。我们设计并使用了一个“人工”数据库进行数据库搜索。总共鉴定了56种液泡膜蛋白，其中70%以上是转运蛋白。砷酸盐处理下，TDT转运蛋白，TerC家族的成员和PDR样蛋白的差异上调。而V-ATPase亚基c、E、G和V-PPase表达下调。此外，本研究中鉴定的液泡膜蛋白为砷载体或通道提供了信息基础，并有助于阐明液泡膜砷转运过程的调控。生物学意义细胞的区室化对砷超积累菌P. vittata中的砷转运蛋白，而已知的参与液泡区室化的砷转运蛋白有限。本论文中，我们获得了P.采用碘克沙醇分步梯度离心法对油蕨叶片的液泡膜蛋白进行了定量分析，并采用TMPP蛋白标记蛋白质组技术对油蕨叶片液泡膜蛋白进行了定量检测。我们的研究结果是对P. vittata，为As载体或通道提供了信息基础。蛋白质组学方法适用于检测液泡膜蛋白的变化，有助于阐明液泡膜运输过程的调控。
Pteris vittata, the first known arsenic hyperaccumulating plant, can accumulate very high concentration arsenic in its aboveground tissues, while low in roots. Previous studies have suggested that arsenic vacuole compartmentalization may play an important role in the arsenic-hyperaccumulation inP. vittata, but the mechanism(s) of arsenic transport to vacuole are largely unknown. We obtained tonoplast isolated from fronds ofP. vittatasporophyte grown under minus and 1 mM arsenate for 3 weeks by iodixanol step gradient centrifugation method, and then used TMPP protein labeling technology followed by liquid chromatography—a linear ion trap-Orbitrap hybrid mass spectrometer analysis for the quantitative detection of proteins. And we designed and used an “artificial” database for database searching. In total, 56 tonoplast proteins were identified; more than 70% of them were transport proteins. Under arsenate treatment, one TDT transporter protein, a member of the TerC family and a PDR-like protein were upregulated differentially. While V-ATPase subunits c, E, and G, and V-PPase, were downregulated. Additionally, the identified tonoplast proteins in our present study provide an informative basis for arsenic carriers or channels and help to clarify the regulation of tonoplast arsenic transport processes inP. vittata.Biological significanceVacuole compartmentalization is crucial to As hyperaccumulatorP. vittata, while there is limited known arsenic transport proteins involved in vacuole compartmentalization. In this paper, we obtained tonoplast ofP. vittatafronds by iodixanol step gradient centrifugation method and then used TMPP protein labeling proteome technology for the quantitative detection of fronds tonoplast proteins. Our findings are the first challenge to the tonoplast proteins data mining ofP. vittatawhich provide an informative basis for As carriers or channels. The proteomic approach in our study is suited for detecting alterations tonoplast protein and help to clarify the regulation of tonoplast transport processes.This article is part of a Special Issue entitled: Proteomics of non-model organisms.
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通讯作者：
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