Analysis of the Arabidopsis mitochondrial proteome
Analysis of the Arabidopsis mitochondrial proteome
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DOI：
10.1104/pp.010387
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发表时间：
2001-12-01
期刊：
影响因子：
7.4
通讯作者：
Leaver, CJ
中科院分区：
文献类型：
--
作者：
Millar, AH;Sweetlove, LJ;Leaver, CJ
目前还不确定编码靶向植物线粒体的蛋白质的完整核基因组，因此植物中线粒体功能的全部范围是未知的。拟南芥细胞培养物线粒体蛋白的二维凝胶分离分析揭示了大约100个丰富的蛋白质和250个低丰度蛋白质。线粒体蛋白质的亚组分的二维凝胶上的比较，提供了信息的可溶性，膜，或膜的整体位置，这种蛋白质：集。共170个蛋白点被切除，胰蛋白酶消化，和基质辅助激光解吸电离/飞行时间质谱光谱获得。使用该数据集，通过搜索翻译的拟南芥基因组数据库鉴定了91种蛋白质。在这一组中，81个定义了基于序列比较的功能。这些功能包括呼吸电子传递、三羧酸循环代谢、氨基酸代谢、蛋白质输入、加工和组装、转录、膜运输和抗氧化防御。总共10个光谱与拟南芥假定的开放阅读框架相匹配，其中没有确定特定的功能。共有64个光谱与识别的开放阅读框架不匹配。使用生物信息学工具来预测亚细胞靶向（TargetP、Psort和MitoProt）的全长推定蛋白质序列的分析揭示了预测的显著变化，并且还缺乏对几种表征的线粒体蛋白质的线粒体靶向预测。
The complete set of nuclear genes that encode proteins targeted to mitochondria in plants is currently undefined and thus the full range of mitochondrial functions in plants is unknown. Analysis of two-dimensional gel separations of Arabidopsis cell culture mitochondrial protein revealed approximately 100 abundant proteins and 250 low-abundance proteins. Comparison of subfractions of mitochondrial protein on two-dimensional gels provided information on the soluble, membrane, or integral membrane locations of this protein :set. A total of 170 protein spots were excised, trypsin-digested, and matrix-assisted laser desorption ionization/time of flight mass spectrometry spectra obtained. Using this dataset, 91 of the proteins were identified by searching translated Arabidopsis genomic databases. Of this set, 81 have defined functions based on sequence comparison. These functions include respiratory electron transport, tricarboxylic acid cycle metabolism, amino acid metabolism, protein import, processing, and assembly, transcription, membrane transport, and antioxidant defense. A total of 10 spectra were matched to Arabidopsis putative open reading frames for which no specific function has been determined. A total of 64 spectra did not match to an identified open reading frame. Analysis of full-length putative protein sequences using bioinformatic tools to predict subcellular targeting (TargetP, Psort, and MitoProt) revealed significant variation in predictions, and also a lack of mitochondrial targeting prediction for several characterized mitochondrial proteins.