Kharel, YugeshAgah, SayehHuang, TaoMendelson, Anna J.Eletu, Oluwafunmilayo T.Barkey-Bircannl, PeterGesualdil, JamesSmith, Jeffrey S.Santos, Webster L.Lynch, Kevin R.2019-06-042019-06-042018-04-19e0192179http://hdl.handle.net/10919/89743Successful medicinal chemistry campaigns to discover and optimize sphingosine kinase inhibitors require a robust assay for screening chemical libraries and for determining rank order potencies. Existing assays for these enzymes are laborious, expensive and/or low throughput. The toxicity of excessive levels of phosphorylated sphingoid bases for the budding yeast, Saccharomyces cerevisiae, affords an assay wherein inhibitors added to the culture media rescue growth in a dose-dependent fashion. Herein, we describe our adaptation of a simple, inexpensive, and high throughput assay for assessing inhibitors of sphingosine kinase types 1 and 2 as well as ceramide kinase and for testing enzymatic activity of sphingosine kinase type 2 mutants. The assay was validated using recombinant enzymes and generally agrees with the rank order of potencies of existing inhibitors.application/pdfenCreative Commons Attribution 4.0 InternationalArticle - Refereedhttps://doi.org/10.1371/journal.pone.0192179134296725281932-6203