TY - JOUR T1 - <em>Saccharomyces cerevisiae</em> as a Platform for Assessing Sphingolipid Lipid Kinase Inhibitors JF - bioRxiv DO - 10.1101/148858 SP - 148858 AU - Yugesh Kharel AU - Sayeh Agah AU - Anna J. Mendelson AU - Oluwafunmilayo T. Eletu AU - James Gesualdi AU - Jeffrey S. Smith AU - Webster L. Santos AU - Kevin R. Lynch Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/06/11/148858.abstract N2 - Successful medicinal chemistry campaigns to discover and improve 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 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 rank order of potencies of existing inhibitors. ER -