Large-scale top-down proteomics of the Arabidopsis thaliana leaf and chloroplast proteomes.
Large-scale top-down proteomics of the Arabidopsis thaliana leaf and chloroplast proteomes.
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作者：
我们提出了一个大规模的自上而下的植物叶片和叶绿体蛋白质组学（TDP）的研究，实现了超过4700个独特的蛋白质组的鉴定。使用毛细管区带电泳结合串联质谱分析离线尺寸排阻色谱馏分，我们确定3198 proteoforms总叶和1836 proteoforms叶绿体，1024和363 proteoforms具有翻译后修饰，分别。毛细管区带电泳的电泳迁移率预测使我们能够验证影响电荷状态的翻译后修饰，如乙酰化和磷酸化。确定的修改包括色氨酸（二）氧化事件的6个叶绿体蛋白，可能代表新的目标单线态氧传感。此外，我们的TDP数据提供了直接的实验证据的N-和C-末端残基的许多成熟的proteoforms从叶绿体，线粒体，内质网，和其他亚细胞定位。有了这些信息，我们建议真正的转运肽裂解位点和正确的亚细胞定位信号预测。这种大规模的分析说明了自上而下的蛋白质型鉴定翻译后修饰和完整序列的能力，这有助于我们了解数百种植物蛋白质的结构和功能。
We present a large-scale top-down proteomics (TDP) study of plant leaf and chloroplast proteins, achieving the identification of over 4700 unique proteoforms. Using capillary zone electrophoresis coupled with tandem mass spectrometry analysis of offline size-exclusion chromatography fractions, we identify 3198 proteoforms for total leaf and 1836 proteoforms for chloroplast, with 1024 and 363 proteoforms having post-translational modifications, respectively. The electrophoretic mobility prediction of capillary zone electrophoresis allowed us to validate post-translational modifications that impact the charge state such as acetylation and phosphorylation. Identified modifications included Trp (di)oxidation events on six chloroplast proteins that may represent novel targets of singlet oxygen sensing. Furthermore, our TDP data provides direct experimental evidence of the N- and C-terminal residues of numerous mature proteoforms from chloroplast, mitochondria, endoplasmic reticulum, and other sub-cellular localizations. With this information, we suggest true transit peptide cleavage sites and correct sub-cellular localization signal predictions. This large-scale analysis illustrates the power of top-down proteoform identification of post-translational modifications and intact sequences that can benefit our understanding of both the structure and function of hundreds of plant proteins.
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影响因子：
14.9
作者：
Heazlewood, Joshua L.;Verboom, Robert E.;Millar, A. Harvey
通讯作者：
Millar, A. Harvey
影响因子：
7.4
作者：
Chen D;Lubeckyj RA;Yang Z;McCool EN;Shen X;Wang Q;Xu T;Sun L
通讯作者：
Sun L
影响因子：
7.4
作者：
Chen B;Brown KA;Lin Z;Ge Y
通讯作者：
Ge Y
影响因子：
5.8
作者：
Kou, Qiang;Xun, Likun;Liu, Xiaowen
通讯作者：
Liu, Xiaowen
影响因子：
4.1
作者：
CIFUENTES, A;POPPE, H
通讯作者：
POPPE, H