Profiling Phosphoproteins of Yeast Mitochondria Reveals a Role of Phosphorylation in Assembly of the ATP Synthase*S

Profiling Phosphoproteins of Yeast Mitochondria Reveals a Role of Phosphorylation in Assembly of the ATP Synthase*S
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酵母线粒体磷蛋白分析揭示磷酸化在 ATP 合酶*S 组装中的作用

DOI:
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发表时间:
2007
影响因子:
7
通讯作者:
C. Meisinger
C. Meisinger
中科院分区:
生物学1区
文献类型:
--
作者:
J. Reinders;K. Wagner;R. Zahedi;Diana Stojanovski;Beate Eyrich;Martin van der Laan;P. Rehling;A. Sickmann;N. Pfanner;C. Meisinger

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线粒体对许多细胞过程至关重要,但线粒体功能的调节仅部分了解。最近的研究表明,线粒体磷蛋白的数量高于预期;然而,可逆磷酸化对线粒体结构和功能的影响仅在少数情况下被定义。因此,从遗传上易处理的生物体中高度纯化的线粒体中确定真实的蛋白质磷酸化位点是至关重要的。酿酒酵母是线粒体功能分析的主要模式生物。我们分离了高纯度的酵母线粒体,并通过不同富集策略和质谱法的组合对磷酸化位点进行了系统分析。我们在48种不同的蛋白质中鉴定了80个磷酸化位点。这些线粒体磷蛋白参与关键的线粒体功能,包括能量代谢、蛋白质生物合成、脂肪酸代谢、代谢物转运和氧化还原调节。通过结合酵母遗传学和体外生化分析,我们发现,磷酸化的丝氨酸残基在亚基g(Atp 20)调节线粒体ATP合酶的二聚化。酵母线粒体磷酸化蛋白质组是揭示可逆性蛋白质磷酸化新作用的丰富来源。
Mitochondria are crucial for numerous cellular processes, yet the regulation of mitochondrial functions is only understood in part. Recent studies indicated that the number of mitochondrial phosphoproteins is higher than expected; however, the effect of reversible phosphorylation on mitochondrial structure and function has only been defined in a few cases. It is thus crucial to determine authentic protein phosphorylation sites from highly purified mitochondria in a genetically tractable organism. The yeast Saccharomyces cerevisiae is a major model organism for the analysis of mitochondrial functions. We isolated highly pure yeast mitochondria and performed a systematic analysis of phosphorylation sites by a combination of different enrichment strategies and mass spectrometry. We identified 80 phosphorylation sites in 48 different proteins. These mitochondrial phosphoproteins are involved in critical mitochondrial functions, including energy metabolism, protein biogenesis, fatty acid metabolism, metabolite transport, and redox regulation. By combining yeast genetics and in vitro biochemical analysis, we found that phosphorylation of a serine residue in subunit g (Atp20) regulates dimerization of the mitochondrial ATP synthase. The authentic phosphoproteome of yeast mitochondria will represent a rich source to uncover novel roles of reversible protein phosphorylation.
DOI: --
发表时间: 1982
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影响因子: --
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