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Identification of Chloroplast Envelope Proteins with Critical Importance for Cold Acclimation.
Plant Physiology ( IF 6.5 ) Pub Date : 2020-01-13 , DOI: 10.1104/pp.19.00947
Oliver Trentmann 1 , Timo Mühlhaus 2 , David Zimmer 2 , Frederik Sommer 3 , Michael Schroda 3 , Ilka Haferkamp 1 , Isabel Keller 1 , Benjamin Pommerrenig 1 , Horst Ekkehard Neuhaus 4
Affiliation  

The ability of plants to withstand cold temperatures relies on their photosynthetic activity. Thus, the chloroplast is of utmost importance for cold acclimation and acquisition of freezing tolerance. During cold acclimation, the properties of the chloroplast change markedly. To provide the most comprehensive view of the protein repertoire of the chloroplast envelope, we analyzed this membrane system in Arabidopsis (Arabidopsis thaliana) using mass spectrometry-based proteomics. Profiling chloroplast envelope membranes was achieved by a cross comparison of protein intensities across the plastid and the enriched membrane fraction under both normal and cold conditions. We used multivariable logistic regression to model the probabilities for the classification of an envelope localization. In total, we identified 38 envelope membrane intrinsic or associated proteins exhibiting altered abundance after cold acclimation. These proteins comprise several solute carriers, such as the ATP/ADP antiporter nucleotide transporter2 (NTT2; substantially increased abundance) or the maltose exporter MEX1 (substantially decreased abundance). Remarkably, analysis of the frost recovery of ntt loss-of-function and mex1 overexpressor mutants confirmed that the comparative proteome is well suited to identify key factors involved in cold acclimation and acquisition of freezing tolerance. Moreover, for proteins with known physiological function, we propose scenarios explaining their possible roles in cold acclimation. Furthermore, spatial proteomics introduces an additional layer of complexity and enables the identification of proteins differentially localized at the envelope membrane under the changing environmental regime.

中文翻译:

叶绿体包膜蛋白的鉴定对于冷驯化至关重要。

植物承受寒冷温度的能力依赖于它们的光合作用活动。因此,叶绿体对于冷驯化和获得冷冻耐受性至关重要。在冷驯化过程中,叶绿体的特性发生显着变化。为了提供叶绿体包膜蛋白质库的最全面视图,我们使用基于质谱的蛋白质组学分析了拟南芥(Arabidopsis thaliana)中的这种膜系统。通过在正常和寒冷条件下交叉比较质体和富集膜部分的蛋白质强度来对叶绿体包膜进行分析。我们使用多变量逻辑回归对包络定位的分类概率进行建模。总共,我们鉴定了 38 种包膜内在蛋白或相关蛋白,它们在冷驯化后表现出丰度改变。这些蛋白质包含几种溶质载体,例如 ATP/ADP 逆向转运蛋白核苷酸转运蛋白 2(NTT2;丰度大幅增加)或麦芽糖输出蛋白 MEX1(丰度大幅下降)。值得注意的是,对 ntt 功能丧失和 mex1 过表达突变体的霜冻恢复分析证实,比较蛋白质组非常适合识别冷驯化和获得冷冻耐受性中涉及的关键因素。此外,对于具有已知生理功能的蛋白质,我们提出了解释它们在冷适应中可能发挥的作用的方案。此外,空间蛋白质组学引入了额外的复杂性,并能够在不断变化的环境条件下识别差异定位在包膜上的蛋白质。
更新日期:2020-03-03
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