Mass spectrometry analysis of proteome-wide proteolytic post-translational degradation of proteins
Mass spectrometry analysis of proteome-wide proteolytic post-translational degradation of proteins
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DOI：
10.1021/ac800077w
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发表时间：
2008-08-01
影响因子：
7.4
通讯作者：
Smith, Richard D.
中科院分区：
文献类型：
--
作者：
Shen, Yufeng;Hixson, Kim K.;Smith, Richard D.
蛋白质降解是细胞正常功能和生命周期的重要组成部分。在这里，我们通过基于测序的串联质谱仪检测蛋白质在胞内降解产生的中间肽，在蛋白质组范围内研究酿酒酵母细胞中蛋白质的降解。通过追踪检测到的与1100个相似的多肽及其与200个蛋白质底物来源相似的来源，我们获得了对酵母细胞中蛋白质组范围内蛋白质选择性降解的新见解的证据。这一证据表明，酵母细胞质是降解具有生化和几何特性的蛋白质的最大池，而在所研究的条件下，酵母核似乎是一个蛋白质分解惰性细胞器。酵母V-ATPase亚基在分解过程中似乎被降解，作为前体、运输载体和GATE的酵母线粒体蛋白优先被降解。根据我们的观察，泛素化对于酵母细胞质调节蛋白和酶蛋白的蛋白酶体降解可能是不必要的。这项研究表明，细胞内肽是直接探索蛋白质降解相关分子机制和细胞生物学过程的信息靶点。
Protein proteolytic degradation is an essential component to proper cell function and its life cycle. Here, we study the protein degradation in yeast Saccharomyces cerevisiae cells on a proteome-wide scale by detection of the intermediate peptides produced from the intracellular degradation of proteins using sequencing-based tandem mass spectrometry. By tracing the detected similar to 1100 peptides and their similar to 200 protein-substrate origins we obtain evidence for new insights into the proteome-wide protein-selective degradation in yeast cells. This evidence shows that the yeast cytoplasm is the largest pool for the degradation of proteins with both biochemical and geometric specificities, whereas the yeast nucleus seems to be a proteolysis-inert organelle tinder the condition studied. Yeast V-ATPase subunits appear to be degraded during their disassembly, and yeast mitochondrial proteins functioning as precursors, transport carriers, and gates are preferentially degraded. Ubiquitylation may be unnecessary for the proteasomal degradation of yeast cytoplasmic regulatory and enzyme proteins according to our observations. This study shows that the intracellular peptides are informational targets for directly probing the protein degradation-involved molecular mechanisms and cell biology processes.