Comparison of the Proteomes and Phosphoproteomes of S. cerevisiae Cells Harvested with Different Strategies.
Comparison of the Proteomes and Phosphoproteomes of S. cerevisiae Cells Harvested with Different Strategies.
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DOI：
10.3390/proteomes11040028
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发表时间：
2023-09-27
期刊：
影响因子：
3.3
通讯作者：
Paulo JA
中科院分区：
文献类型：
--
作者：
Rossio V;Paulo JA
芽殖酵母酿酒酵母是一个功能强大的模型系统，被广泛用于研究许多细胞过程。收获酵母细胞是几乎所有实验程序的第一步。在这里，酵母细胞从其生长培养基中分离，收集，并用于连续的实验或分析。两种最常用的收获S.酿酒酵母是离心和过滤。了解离心和过滤是否以及如何影响酵母生理学对于下游数据解释至关重要。在这里，我们概况和比较的蛋白质组和磷酸化蛋白质组，使用同量异位素标记为基础的定量质谱，三种常用的方法来收获S。酿酒酵母细胞：低速离心、高速离心和过滤。我们的数据表明，虽然蛋白质组在测试条件下是稳定的，但离心和过滤之间的数百个磷酸化事件是不同的。我们的分析表明，在我们的实验条件下，过滤可能会导致细胞壁和渗透应力在更高的水平相比，离心，这意味着收获方法的具体应力。因此，考虑到特定应力的基础激活水平在某些收获条件下可能不同，这是实验设计的一个重要但经常被忽视的方面。
The budding yeast Saccharomyces cerevisiae is a powerful model system that is widely used to investigate many cellular processes. The harvesting of yeast cells is the first step in almost every experimental procedure. Here, yeast cells are isolated from their growth medium, collected, and used for successive experiments or analysis. The two most common methods to harvest S. cerevisiae are centrifugation and filtration. Understanding if and how centrifugation and filtration affect yeast physiology is essential with respect to downstream data interpretation. Here, we profile and compare the proteomes and the phosphoproteomes, using isobaric label-based quantitative mass spectrometry, of three common methods used to harvest S. cerevisiae cells: low-speed centrifugation, high-speed centrifugation, and filtration. Our data suggest that, while the proteome was stable across the tested conditions, hundreds of phosphorylation events were different between centrifugation and filtration. Our analysis shows that, under our experimental conditions, filtration may cause both cell wall and osmotic stress at higher levels compared to centrifugation, implying harvesting-method-specific stresses. Thus, considering that the basal activation levels of specific stresses may differ under certain harvesting conditions is an important, but often overlooked, aspect of experimental design.
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影响因子：
64.5
作者：
Huttlin EL;Jedrychowski MP;Elias JE;Goswami T;Rad R;Beausoleil SA;Villén J;Haas W;Sowa ME;Gygi SP
通讯作者：
Gygi SP
影响因子：
14.9
作者：
Hornbeck PV;Kornhauser JM;Tkachev S;Zhang B;Skrzypek E;Murray B;Latham V;Sullivan M
通讯作者：
Sullivan M
影响因子：
3.3
作者：
Paulo, Joao A.;O'Connell, Jeremy D.;Gygi, Steven P.
通讯作者：
Gygi, Steven P.
影响因子：
5.6
作者：
Vanderwaeren, Laura;Dok, Ruveyda;Voordeckers, Karin;Nuyts, Sandra;Verstrepen, Kevin J.
通讯作者：
Verstrepen, Kevin J.
影响因子：
3.3
作者：
Levin, David E.
通讯作者：
Levin, David E.