Global analysis of protein phosphorylation in yeast
Global analysis of protein phosphorylation in yeast
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DOI：
10.1038/nature04187
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发表时间：
2005-12-01
期刊：
影响因子：
64.8
通讯作者：
Snyder, M
中科院分区：
文献类型：
--
作者：
Ptacek, J;Devgan, G;Snyder, M
据估计，蛋白质磷酸化影响30%的蛋白质组，是一种控制许多基本细胞过程的主要调节机制（1 - 3）。直到最近，我们在全球范围内对蛋白质磷酸化的生化理解还极其有限；只有一半的酵母激酶已知其体内底物，而且已知其磷酸化激酶的磷蛋白不到160种。在此，我们利用蛋白质组芯片技术（4）描述了大多数酵母蛋白激酶（5）所识别的体外底物：我们确定了涉及1325种不同蛋白质的4000多个磷酸化事件。这些底物代表了不同生化功能和细胞作用的广泛范围。每种蛋白激酶都能识别不同的底物组，包括蛋白激酶A家族中密切相关的激酶以及仅在细胞周期蛋白亚基上有所不同的四种细胞周期蛋白依赖性激酶。尽管许多底物与它们的磷酸化激酶位于同一细胞区室或属于同一功能类别，但也有许多并非如此，这表明一些激酶可能具有新的作用。此外，将磷酸化结果与蛋白质 - 蛋白质相互作用（6 - 10）以及转录因子结合数据（11，12）相结合，揭示了新的调节模块。我们的磷酸化结果已被整合为酵母的第一代磷酸化图谱。由于许多酵母蛋白质和通路是保守的，这些结果将为理解许多真核生物中蛋白质磷酸化的机制和作用提供见解。
Protein phosphorylation is estimated to affect 30% of the proteome and is a major regulatory mechanism that controls many basic cellular processes(1-3). Until recently, our biochemical understanding of protein phosphorylation on a global scale has been extremely limited; only one half of the yeast kinases have known in vivo substrates and the phosphorylating kinase is known for less than 160 phosphoproteins. Here we describe, with the use of proteome chip technology(4), the in vitro substrates recognized by most yeast protein kinases(5): we identified over 4,000 phosphorylation events involving 1,325 different proteins. These substrates represent a broad spectrum of different biochemical functions and cellular roles. Distinct sets of substrates were recognized by each protein kinase, including closely related kinases of the protein kinase A family and four cyclin- dependent kinases that vary only in their cyclin subunits. Although many substrates reside in the same cellular compartment or belong to the same functional category as their phosphorylating kinase, many others do not, indicating possible new roles for several kinases. Furthermore, integration of the phosphorylation results with protein - protein interaction(6-10) and transcription factor binding data(11,12) revealed novel regulatory modules. Our phosphorylation results have been assembled into a first- generation phosphorylation map for yeast. Because many yeast proteins and pathways are conserved, these results will provide insights into the mechanisms and roles of protein phosphorylation in many eukaryotes.