Journal
JOURNAL OF BIOMOLECULAR SCREENING
Volume 13, Issue 9, Pages 906-911Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057108324498
Keywords
PI 3-kinase p110 alpha; liposome; lipid extraction; high-throughput screening
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The signaling pathways involving lipid kinase class I phosphatidylinositol 3-kinases (PI 3-kinases) regulate cell growth, proliferation, and survival. Class I PI 3-kinases catalyze the conversion of PI (4.5)P-2 to PI (3.4,5)P-3, which acts as a lipid second messenger to activate mitogenic signaling cascades. Recently, p110 alpha class IA PI 3-kinase, was found to be mutated frequently in many human cancers. Therefore. it is increasingly studied as an anticancer drug target. Traditionally, PI 3-kinase activities have been studied using liposome substrates. This method, however, is hampered significantly by the labor-intensive manual lipid extraction followed by a low-throughput thin-layer chromatography analysis. The authors describe a high-throughput liposome substrate-based assay based on an automated lipid extraction method that allows them to study PI 3-kinase enzyme mechanism and quantitatively measure inhibitor activity using liposome substrates in a high-throughput mode. This improved assay format can easily be extended to Study other classes of phosphoinositide lipid kinases. (Journal of Biomolecular Screening 2008:906-911)
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