Saccharomyces cerevisiae is an established model organism with a well characterized genome. However, this model presents a unique problem due to a very resistant cell wall which develops in the late stationary phase resulting in sub-optimal extraction of proteins from such cells using majority of the cell lysis protocols. In this study, several methods from the literature with modifications thereof for lysis of S. cerevisiae cells were analyzed for their suitability for redox proteomics and biological activity studies of both exponential and late stationary phase cultures. The protocols applied are glass bead lysis, sonication, their combinations, alkali extraction, hot-SDS extraction methods and their modifications. The glass bead lysis method showed low yield but could be convenient in cases where in vitro processing steps post extraction is required or if only hydrophilic proteins are of interest. Hot-SDS and alkali extraction protocols yielded higher amount of proteins and these methods are potentially suitable for Western blotting and redox proteomic studies but allow no post-processing treatment(s) on the extracts which may be required for aging- and oxidative stress-related or other studies.

中文翻译：

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在不同生长阶段从酿酒酵母中提取蛋白质。

酿酒酵母是具有良好表征的基因组的成熟模型生物。然而，由于在晚期固定相中形成的极高抗性的细胞壁导致使用大多数细胞裂解方案从此类细胞中提取蛋白质的效果欠佳，因此该模型提出了一个独特的问题。在这项研究中，分析了文献中几种经修饰的酿酒酵母细胞裂解方法，它们是否适用于指数和后期固定相培养的氧化还原蛋白质组学和生物学活性研究。应用的协议是玻璃珠裂解，超声处理，它们的组合，碱提取，热SDS提取方法及其修改方法。玻璃珠裂解法显示出低产率，但是在需要提取后的体外加工步骤或仅对亲水性蛋白质感兴趣的情况下可能很方便。热SDS和碱提取方案产生的蛋白质量更高，这些方法可能适用于蛋白质印迹和氧化还原蛋白质组学研究，但不允许对提取物进行后期处理，而老化和氧化应激相关可能需要这些提取物或其他研究。