1. Saccharomyces cerevisiae as a platform for assessing sphingolipid lipid kinase inhibitors.
- Author
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Kharel, Yugesh, Agah, Sayeh, Huang, Tao, Mendelson, Anna J., Eletu, Oluwafunmilayo T., Barkey-Bircann, Peter, Gesualdi, James, Smith, Jeffrey S., Santos, Webster L., and Lynch, Kevin R.
- Subjects
SACCHAROMYCES cerevisiae ,SPHINGOLIPIDS ,KINASE inhibitors ,CHEMICAL libraries ,PHARMACEUTICAL chemistry - Abstract
Successful 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. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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