A fluorescent-based high-throughput method to quantitatively measuring PSS1-mediated phosphatidylserine synthesis
摘要
With phosphatidylcholine and L-serine, phosphatidylserine synthase 1 (PSS1) synthesizes phosphatidylserine, a phospholipid that is a key component in cell membranes. Here, we provide a step-by-step protocol examining the enzymatic acidity of PSS1 in vitro. This in vitro assay quantitatively measures Choline, a byproduct generated in the biochemical reaction, featuring simple operation, high sensitivity and low cost without the requirement for radioactive labeling. It enables rapid evaluation of PSS1 activity and is applicable to high-throughput screening of PSS1 inhibitors.
参考文献
1. Zhang, F. and G. Du, Dysregulated lipid metabolism in cancer. World J Biol Chem, 2012. 3(8): p. 167–74.
2. Gupta, A., D. Das, and R. Taneja, Targeting Dysregulated Lipid Metabolism in Cancer with Pharmacological Inhibitors, in Cancers (Basel). 2024: Switzerland.
3. Leventis, P.A. and S. Grinstein, The distribution and function of phosphatidylserine in cellular membranes. Annu Rev Biophys, 2010. 39: p. 407–27.
4. Tomohiro, S., et al., Purification and characterization of human phosphatidylserine synthases 1 and 2, in Biochem J. 2009: England. p. 421–9.
5. Yoshihama, Y., et al., Potent and Selective PTDSS1 Inhibitors Induce Collateral Lethality in Cancers with PTDSS2 Deletion, in Cancer Res. 2022: United States. p. 4031–4043.
6. Omi, J., et al., Phosphatidylserine synthesis controls oncogenic B cell receptor signaling in B cell lymphoma, in J Cell Biol. 2024: United States.
7. Suzuki, K., et al., Efficient Multikilogram-Scale Synthesis of PTDSS1 Inhibitor: Development of a Practical and Scalable Optical Resolution Method for Chiral 2,3-Pyrrolidinedione. Organic Process Research & Development, 2024. 28(6): p. 2296–2308.
指标
DOI：
Submission ID：
下载次数
已发布
如何引用
下载引用
利益冲突声明
作者声明无任何需要披露的利益冲突。
Copyright
本预印本的版权持有者为作者/资助方。
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.