Saccharomyces cerevisiae as a platform for assessing sphingolipid kinase inhibitors

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 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 rank order of potencies of existing inhibitors.

Download data

• Downloaded 558 times
• Download rankings, all-time:
  • Site-wide: 40,316
  • In biochemistry: 1,036
• Year to date:
  • Site-wide: 69,857
• Since beginning of last month:
  • Site-wide: 47,499

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide