A fluorescent-based high-throughput method to quantitatively measuring PSS1-mediated phosphatidylserine synthesis
Abstract
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.
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The authors declare no competing interests to disclose.
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