Graduate School of Biosphere Science, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima 739-8528, Japan.
Graduate School of Integrated Sciences for Life, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima 739-8528, Japan.
STAR Protoc. 2021 Apr 10;2(2):100412. doi: 10.1016/j.xpro.2021.100412. eCollection 2021 Jun 18.
Sphingolipid biosynthesis occurs in both the endoplasmic reticulum (ER) and the Golgi apparatus. Ceramide synthesized in the ER is transported to the Golgi and incorporated into complex sphingolipids. Here, we present a step-by-step protocol to analyze sphingolipid metabolism in budding yeast. Ceramide and inositolphosphorylceramide (IPC) are classes of sphingolipids present in yeast and are metabolically labeled with radioactive precursors. This protocol for metabolic labeling can be used to investigate ceramide transport in an environment. For complete details on the use and execution of this protocol, please refer to Ikeda et al. (2020).
鞘脂类生物合成发生在内质网（ER）和高尔基体中。在内质网中合成的神经酰胺被运输到高尔基体并掺入到复合鞘脂类中。在这里，我们提供了一个逐步的方案来分析出芽酵母中的鞘脂代谢。神经酰胺和肌醇磷酸神经酰胺（IPC）是酵母中存在的两类鞘脂，它们可以用放射性前体进行代谢标记。这个代谢标记的方案可以用于研究环境中的神经酰胺运输。有关此方案的使用和执行的完整详细信息，请参阅 Ikeda 等人。（2020 年）。
