QUEEN-based Spatiotemporal ATP Imaging in Budding and Fission Yeast
QUEEN-based Spatiotemporal ATP Imaging in Budding and Fission Yeast
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DOI：
10.21769/bioprotoc.3320
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发表时间：
2019-08-05
期刊：
影响因子：
0.8
通讯作者：
Takaine, Masak
中科院分区：
文献类型：
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作者：
Takaine, Masak
酵母为研究真核细胞的代谢和生物能量学提供了一个特殊的模型。在众多的代谢物中，三磷酸腺苷（ATP）是所有生物体所必需的主要代谢物。因此，更清楚地了解活酵母细胞中的ATP动力学对于破译细胞能量代谢是重要的。然而，目前可用于测量ATP的方法，包括生化分析和ATP指示剂，都不适合在单细胞水平上仔细检查酵母细胞中的ATP浓度。使用最近开发的ATP生物传感器QUEEN，这是适合于酵母和细菌，本文描述了一个协议，以可视化ATP浓度活芽殖和分裂酵母细胞。这种简单的方法使各种酵母突变体中ATP动力学的检查变得简单可靠，从而为细胞能量代谢提供了新的分子见解。
Yeasts have provided an exceptional model for studying metabolism and bioenergetics in eukaryotic cells. Among numerous metabolites, adenosine triphosphate (ATP) is a major metabolite that is essential for all living organisms. Therefore, a clearer understanding of ATP dynamics in living yeast cells is important for deciphering cellular energy metabolism. However, none of the methods currently available to measure ATP, including biochemical analyses and ATP indicators, have been suitable for close examinations of ATP concentrations in yeast cells at the single cell level. Using the recently developed ATP biosensor QUEEN, which is suitable for yeasts and bacteria, a protocol was described herein to visualize ATP concentrations in living budding and fission yeast cells. This simple method enables the easy and reliable examination of ATP dynamics in various yeast mutants, thereby providing novel molecular insights into cellular energy metabolism.