Shen Hongling, He Zhenyan, Yan Huili, Xing Zenan, Chen Yanshan, Xu Wenxiu, Xu Wenzhong, Ma Mi
Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
J Proteomics. 2014 Jun 13;105:46-57. doi: 10.1016/j.jprot.2014.01.029. Epub 2014 Feb 4.
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 in P. vittata, but the mechanism(s) of arsenic transport to vacuole are largely unknown. We obtained tonoplast isolated from fronds of P. vittata sporophyte grown under minus and 1mM arsenate for 3weeks 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 in P. vittata.
Vacuole compartmentalization is crucial to As hyperaccumulator P. vittata, while there is limited known arsenic transport proteins involved in vacuole compartmentalization. In this paper, we obtained tonoplast of P. vittata fronds 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 of P. vittata which 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.
蜈蚣草是已知的第一种砷超富集植物，其地上组织能积累高浓度的砷，而根部的砷含量较低。先前的研究表明，砷在液泡中的区室化作用可能在蜈蚣草的砷超积累过程中发挥重要作用，但砷转运至液泡的机制在很大程度上尚不清楚。我们通过碘克沙醇梯度离心法，从在 - 条件下和1mM砷酸盐中生长3周的蜈蚣草孢子体的叶片中分离得到液泡膜，然后使用TMPP蛋白质标记技术，随后通过液相色谱 - 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。并且我们设计并使用了一个“人工”数据库进行数据库搜索。总共鉴定出56种液泡膜蛋白；其中70%以上是转运蛋白。在砷酸盐处理下，一种TDT转运蛋白、TerC家族的一个成员和一种类PDR蛋白被差异上调。而V - ATPase亚基c、E和G以及V - PPase则被下调。此外，我们在本研究中鉴定出的液泡膜蛋白为砷载体或通道提供了信息基础，并有助于阐明蜈蚣草液泡膜砷转运过程的调控机制。
液泡区室化对于砷超富集植物蜈蚣草至关重要，而参与液泡区室化的已知砷转运蛋白有限。在本文中，我们通过碘克沙醇梯度离心法获得了蜈蚣草叶片的液泡膜，然后使用TMPP蛋白质标记蛋白质组技术对叶片液泡膜蛋白进行定量检测。我们的发现是对蜈蚣草液泡膜蛋白数据挖掘的首次挑战，为砷载体或通道提供了信息基础。我们研究中的蛋白质组学方法适用于检测液泡膜蛋白的变化，并有助于阐明液泡膜转运过程的调控机制。本文是名为：非模式生物蛋白质组学的特刊的一部分。
