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首页> 外文期刊>Journal of proteomics >Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S-cerevisiae proteins across 10 carbon sources
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Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S-cerevisiae proteins across 10 carbon sources

机译:基于定量的质谱 - 基于质量光谱的多路复用比较了10个碳源的5000 s-cerevisiae蛋白的丰度

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The budding yeast Saccharomyces cerevisiae is a model system for investigating biological processes. Cellular events are known to be dysregulated due to shifts in carbon sources. However, the comprehensive proteomic alterations thereof have not been fully investigated. Here we examined proteomic alterations in S. cerevisiae due to the adaptation of yeast from glucose to nine different carbon sources - maltose, trehalose, fructose, sucrose, glycerol, acetate, pyruvate, lactic acid, and oleate. Isobaric tag-based mass spectrometry techniques are at the forefront of global proteomic investigations. As such, we used a TMT10-plex strategy to study multiple growth conditions in a single experiment. The SPS-MS3 method on an Orbitrap Fusion Lumos mass spectrometer enabled the quantification of over 5000 yeast proteins across ten carbon sources at a 1% protein-level FDR. On average, the proteomes of yeast cultured in fructose and sucrose deviated the least from those cultured in glucose. As expected, gene ontology classification revealed the major alteration in protein abundances occurred in metabolic pathways and mitochondrial proteins. Our protocol lays the groundwork for further investigation of carbon source induced protein alterations. Additionally, these data offer a hypothesis-generating resource for future studies aiming to investigate both characterized and uncharacterized genes.
机译:萌芽酵母酿酒酵母是用于调查生物过程的模型系统。已知蜂窝事件由于碳源的偏移而感到困扰。然而,综合蛋白质组学改变尚未完全研究。在这里,我们通过从葡萄糖的适应从葡萄糖到九种不同的碳源 - 麦芽糖,海藻糖,果糖,蔗糖,甘油,乙酸酯,丙酮酸,乳酸和油酸酯,检查了酿酒酵母的蛋白质组学改变。基于ISObaric标签的质谱技术是全球蛋白质组学研究的最前沿。因此,我们使用了TMT10-PLEX策略来研究单一实验中的多种生长条件。在玻璃岩融合钻孔质谱仪上的SPS-MS3方法使得在1%蛋白水平FDR处的10个碳源上量化超过5000酵e蛋白。平均而言,在果糖和蔗糖中培养的酵母蛋白质偏离葡萄糖中培养的那些。正如预期的那样,基因本体类别分类揭示了代谢途径和线粒体蛋白发生的蛋白质丰度的主要变化。我们的协议为进一步调查碳源诱导蛋白质改变奠定了基础。另外,这些数据提供了一个假设产生的资源,用于将来的研究,旨在调查表征和无表征基因。

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