PT  - JOURNAL ARTICLE
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
TI  - <em>Saccharomyces cerevisiae</em> as a Platform for Assessing Sphingolipid Lipid Kinase Inhibitors
AID  - 10.1101/148858
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 148858
4099  - http://biorxiv.org/content/early/2017/06/11/148858.short
4100  - http://biorxiv.org/content/early/2017/06/11/148858.full
AB  - 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.