| Signaling Assays for Detection of Human G-protein-coupled Receptors in Yeast
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Author:
Date:
2014-08-20
[Abstract] G-protein-coupled receptors (GPCRs) are the largest group of cell-surface proteins and are major molecular targets for drug development. The protocol described herein is for the detection of human GPCR signaling in the yeast Saccharomyces cerevisiae. Using Zoanthus sp. green fluorescent protein (ZsGreen) as the reporter, engineered yeast cells expressing human GPCRs emit strong fluorescence in response to stimuli leading to receptor signal activation. This assay method would allow screening for agonistic ligands and critical mutations required for human GPCR signaling.
[摘要] G蛋白偶联受体（GPCR）是最大的细胞表面蛋白质组，是药物开发的主要分子靶标。 本文所述的方案用于在酵母酿酒酵母中检测人GPCR信号传导。 使用 Zoanthus 绿色荧光蛋白（ZsGreen ）作为报告物，表达人GPCR的工程化酵母细胞响应于导致受体信号活化的刺激而发射强荧光。 该测定方法将允许筛选激动剂配体和人GPCR信号传导所需的关键突变。
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| Metabolic Labeling of Yeast Sphingolipids with Radioactive D-erythro-[4,5-3H]dihydrosphingosine
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Author:
Date:
2013-08-20
[Abstract] Yeast sphingolipids can be metabolically labeled with exogenously added radioactive precursors. Here we describe a method to label ceramides and complex sphingolipids of Saccharomyces cerevisiae with a radioactive ceramide precursor, D-erythro-[4, 5-3H]dihydrosphingosine. This protocol is used to study the biosynthesis, transport and metabolism of sphingolipids in yeast.
[摘要] 酵母鞘脂可以用外源添加的放射性前体代谢标记。 在这里我们描述了用放射性神经酰胺前体D-赤式 - [4,5] 3 H]二氢鞘氨醇标记神经酰胺和酿酒酵母的复杂神经鞘脂的方法。 该方案用于研究酵母中鞘脂的生物合成，转运和代谢。
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