A robust screen for inhibitors and enhancers of phosphoinositide-3 kinase (PI3K) activities by ratiometric fluorescence superquenching

Journal of Biomolecular Screening

Volumn 11, Issue 4, 2006, Pages 413-422

  Stankewicz, Casey ^a   Rininsland, Frauke H ^a  

^a QTL Biosystems   (United States)

Author keywords

Cancer; Diabetes; High throughput screening; Phosphoinositide 3 kinases; Superquenching

Indexed keywords

DYE; ENZYME; ENZYME ACTIVATOR; METAL ION; PHOSPHATIDYLINOSITOL 3 KINASE; PHOSPHATIDYLINOSITOL 3 KINASE INHIBITOR; PHOSPHOINOSITIDE 3 KINASE ALPHA; PHOSPHOINOSITIDE 3 KINASE BETA; PHOSPHOINOSITIDE 3 KINASE DELTA; PHOSPHOINOSITIDE 3 KINASE GAMMA; POLYMER; UNCLASSIFIED DRUG;

ARTICLE; AUTOFLUORESCENCE; ENERGY TRANSFER; ENZYME ACTIVITY; ENZYME ASSAY; FLUORESCENCE; HIGH THROUGHPUT SCREENING; PHOSPHORYLATION; PRIORITY JOURNAL;

1-PHOSPHATIDYLINOSITOL 3-KINASE; ANDROSTADIENES; CHROMONES; DRUG EVALUATION, PRECLINICAL; ENZYME INHIBITORS; ISOENZYMES; MORPHOLINES; SPECTROMETRY, FLUORESCENCE;

EID: 33744935308     PISSN: 10870571     EISSN: 1552454X     Source Type: Journal    
DOI: 10.1177/1087057106286402     Document Type: Article

References (15)

1. Cohen P: The origins of protein phosphorylation. Nat Cell Biol 2002;4:127-130.
2. Vanhaesebroeck B, Waterfield MD: Signaling by distinct classes of phosphoinositide 3-kinases. Exp Cell Res 1999;1:239-254.
3. Fuchikami K, Togame H, Sagara A, Satoh T, Gantner F, Bacon KB, et al: A versatile high-throughput screen for inhibitors of lipid kinase activity: development of an immobilized phospholipid plate assay for phosphoinositide 3-kinase gamma. J Biomol Screen 2002;7:441-450.
4. Drees BE, Weipert A, Hudson H, Ferguson CG, Chakravarty L, Prestwich GD: Competitive fluorescence polarization assays for the detection of phosphoinositide kinase and phosphatase activity. Comb Chem High Throughput Screen 2003;6:321-330.
5. Rodems SM, Hamman BD, Lin C, Zhao J, Shall S, Heidary D, et al: A FRET-based assay platform for ultra-high density drug screening of protein kinases and phosphatases. Assay Drug Dev Technol 2002;1:9-19.
6. Gray A, Olsson H, Batty IH, Priganica L, Downes R: Nonradioactive methods for the assay of phosphoinositide 3-kinases and phosphoinositide phosphatases and selective detection of signaling lipids in cell and tissue extracts Anal Biochem 2003;313:234-245.
7. Rininsland F, Xia W, Wittenburg S, Shi X, Stankewicz C, Achyuthan K, et al: Metal ion-mediated polymer superquenching for highly sensitive detection of kinase and phosphatase activities. Proc Natl Acad Sci USA 2004;101:15295-15300.
8. Chen L, McBranch DW, Wang HL, Helgeson R, Wudl F, Whitten DG: Highly sensitive biological and chemical sensors based on reversible fluorescence quenching in a conjugated polymer. Proc Natl Acad Sci USA 1999;96:12287-12289.
9. Rininsland F, Stankewicz C, Weatherford W, McBranch DW: High-throughput kinase assays with protein substrates using fluorescent polymer superquenching. BMC Biotechnol 2005;5:16.
10. Kushon S, Bradford K, Marin V, Suhrada C, Armitage BA, McBranch DW, et al: Detection of single nucleotide mismatches via fluorescent polymer superquenching. Langmuir 2003;19:6456-6464.
11. Vedvik KL, Eliason HC, Hoffman RL, Gibson JR, Kupcho KR, Somberg RL, et al: Overcoming compound interference in fluorescence polarization-based kinase assays using far-red tracers. Assay Drug Dev Technol 2004;2:193-203.
12. Zhang JH, Chung TD, Oldenburg KR: A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 1993;4:67-73.
13. Powis G, Bonjouklian R, Berggren MM, Gallegos A, Abraham R, Ashendel C, et al: Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3- kinase. Cancer Res 1994;54:2419-2423.
14. Vlahos CJ, Matter WF, Hui KY, Brown RF: A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem 1994;269:5241-5248.
15. Matter WF, Brown RF, Vlahos CJ: The inhibition of phosphatidylinositol 3-kinase by quercetin and analogs. Biochem Biophys Res Commun 1992;185:624-631.