Rapid Cryopurification of the Yeast Mitochondrial Ribosome.
Rapid Cryopurification of the Yeast Mitochondrial Ribosome.
复制标题
酵母线粒体核糖体的快速冷冻纯化。
DOI：
10.1007/978-1-0716-3171-3_9
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Barrientos,Antoni
中科院分区：
文献类型：
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作者：
Pang,HongWeng;Barrientos,Antoni
低温研磨或低温研磨涉及使用液氮来降低生物材料和/或研磨过程的温度。当应用于亚细胞或亚细胞器结构和过程的研究时，它允许从冷冻在所选择的生理状态下的全细胞中快速提取它们。这种方法已被证明是有用的酵母线粒体核糖体的研究。冷冻研磨100 mL酵母培养物后，可在天然条件下免疫沉淀并纯化方便标记的线粒体核糖体。这些核糖体适用于下游方法的应用。这些包括线粒体蛋白质组分析，以分析线粒体翻译组或质谱分析，以评估正常生长条件下或应激下的线粒体蛋白质组，如本方法所述。
Cryogenic milling, or cryomilling, involves the use of liquid nitrogen to lower the temperature of the biological material and/or the milling process. When applied to the study of subcellular or suborganellar structures and processes, it allows for their rapid extraction from whole cells frozen in the physiological state of choice. This approach has proven to be useful for the study of yeast mitochondrial ribosomes. Following cryomilling of 100 mL of yeast culture, conveniently tagged mitochondrial ribosomes can be immunoprecipitated and purified in native conditions. These ribosomes are suitable for the application of downstream approaches. These include mitoribosome profiling to analyze the mitochondrial translatome or mass spectrometry analyses to assess the mitoribosome proteome in normal growth conditions or under stress, as described in this method.