The mitochondrial complexome of Arabidopsis thaliana
The mitochondrial complexome of Arabidopsis thaliana
复制标题
DOI：
10.1111/tpj.13448
复制
发表时间：
2017-03
期刊：
影响因子：
--
通讯作者：
Jennifer Senkler;Michael Senkler;Holger Eubel;Tatjana M. Hildebrandt;Christian Lengwenus;P. Schertl;M. Sc
中科院分区：
文献类型：
--
作者：
Jennifer Senkler;Michael Senkler;Holger Eubel;Tatjana M. Hildebrandt;Christian Lengwenus;P. Schertl;M. Sc
线粒体是细胞新陈代谢和能量转换的中心。在植物中，它们还通过额外的成分和功能灵活性实现光合作用。这些过程中的大部分依赖于将单个蛋白质组装成更大的蛋白质复合体，其中一些作为大分子机器运行。植物线粒体蛋白质复合体的组成和功能以及蛋白质与蛋白质的相互作用引起了人们的浓厚兴趣，但所使用的实验方法通常存在选择性或偏差。在这里，我们提出了一个模式植物拟南芥叶片线粒体的复杂组谱分析，以系统地表征蛋白质组装。用1D Blue Natural(BN)PAGE分离纯化的细胞器提取物，将得到的凝胶通道切成70片(复合体部分)，用无标记的定量枪炮蛋白质组学鉴定每片中的蛋白质。总体而言，共鉴定出1359种独特的蛋白质，平均每个存在于17个复杂组份中。沿着BN凝胶通道的蛋白质的定量图谱通过相似性对齐，使我们能够可视化蛋白质组装。这些数据允许重新注释OXPHOS复合体的亚基组成，鉴定OXPHOS复合体的组装中间体和替代呼吸氧化还原酶的组装。发现了一些在植物中尚未报道的蛋白质复合体，如530 kDa的TAT复合体、460 kDa和1000 kDa的SAM复合体、150 kDa的钙离子单转运体复合体和几个PPR蛋白质复合体。我们已经建立了一个定制的在线资源(https://complexomemap.de/at_mito_leaves))来存储数据，并允许直接访问和定制数据分析。
Mitochondria are central to cellular metabolism and energy conversion. In plants they also enable photosynthesis through additional components and functional flexibility. A majority of those processes relies on the assembly of individual proteins to larger protein complexes, some of which operate as large molecular machines. There has been a strong interest in the makeup and function of mitochondrial protein complexes and protein-protein interactions in plants, but the experimental approaches used typically suffer from selectivity or bias. Here, we present a complexome profiling analysis for leaf mitochondria of the model plant Arabidopsis thaliana for the systematic characterization of protein assemblies. Purified organelle extracts were separated by 1D Blue native (BN) PAGE, a resulting gel lane was dissected into 70 slices (complexome fractions) and proteins in each slice were identified by label free quantitative shot-gun proteomics. Overall, 1359 unique proteins were identified, which were, on average, present in 17 complexome fractions each. Quantitative profiles of proteins along the BN gel lane were aligned by similarity, allowing us to visualize protein assemblies. The data allow re-annotating the subunit compositions of OXPHOS complexes, identifying assembly intermediates of OXPHOS complexes and assemblies of alternative respiratory oxidoreductases. Several protein complexes were discovered that have not yet been reported in plants, such as a 530 kDa Tat complex, 460 and 1000 kDa SAM complexes, a calcium ion uniporter complex (150 kDa) and several PPR protein complexes. We have set up a tailored online resource (https://complexomemap.de/at_mito_leaves) to deposit the data and to allow straightforward access and custom data analyses.