Molecular Components of Arabidopsis Intact Vacuoles Clarified with Metabolomic and Proteomic Analyses
Molecular Components of Arabidopsis Intact Vacuoles Clarified with Metabolomic and Proteomic Analyses
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DOI：
10.1093/pcp/pcy069
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发表时间：
2018-07-01
影响因子：
4.9
通讯作者：
Mimura, Tetsuro
中科院分区：
文献类型：
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作者：
Ohnishi, Miwa;Anegawa, Aya;Mimura, Tetsuro
我们使用毛细管电泳质谱 (CE-MS)、傅里叶变换离子回旋共振 (FT-ICR)-MS 和液相色谱 (LC)-MS 分析了从拟南芥悬浮培养细胞中分离的纯完整液泡中所含的代谢物和蛋白质。我们通过 CE-MS 鉴定出 21 种氨基酸和 5 种有机酸是液泡中的主要初级代谢物。此外，我们还鉴定出了少量的 27 种物质，包括众所周知的液泡分子，但也有一些意想不到的物质（例如有机磷酸盐化合物）。使用 FT-ICR-MS 对液泡样品进行非靶向分析表明，有 1,106 个 m/z 峰可以预测 5,090 个分子式，并且我们使用 KNapSAck 数据库注释了这些峰中的 34 个化合物。通过对液泡液进行蛋白质组分析，我们在同一液泡样本中发现了 186 种蛋白质。由于液泡被认为是主要的降解隔室，因此其中许多是水解酶，但我们也发现了各种氧化还原酶和转移酶。讨论了液泡中蛋白质和代谢物之间的关系。
We analyzed the metabolites and proteins contained in pure intact vacuoles isolated from Arabidopsis suspension-cultured cells using capillary electrophoresis-mass spectrometry (CE-MS), Fourier transform-ion cyclotron resonance (FT-ICR)-MS and liquid chromatography (LC)-MS. We identified 21 amino acids and five organic acids as major primary metabolites in the vacuoles with CE-MS. Further, we identified small amounts of 27 substances including well-known vacuolar molecules, but also some unexpected substances (e.g. organic phosphate compounds). Non-target analysis of the vacuolar sample with FT-ICR-MS suggested that there are 1,106 m/z peaks that could predict the 5,090 molecular formulae, and we have annotated 34 compounds in these peaks using the KNapSAck database. By conducting proteomic analysis of vacuolar sap, we found 186 proteins in the same vacuole samples. Since the vacuole is known as a major degradative compartment, many of these were hydrolases, but we also found various oxidoreductases and transferases. The relationships between the proteins and metabolites in the vacuole are discussed.