Document Detail


Kinetic analysis of PI3K reactions with fluorescent PIP2 derivatives.
MedLine Citation:
PMID:  21789487     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Phosphatidylinositol 3-kinase (PI3K) signaling plays important roles in cell differentiation, proliferation, and migration. Increased mutations and expression levels of PI3K are hallmarks for the development of certain cancers. Pharmacological targeting of PI3K activity has also been actively pursued as a novel cancer therapeutic. Consequently, measurement of PI3K activity in different cell types or patient samples holds the promise as being a novel diagnostic tool. However, the direct measurement of cellular PI3K activity has been a challenging task. We report here the characterization of two fluorescent PIP(2) derivatives as reporters for PI3K enzymatic activity. The reporters are efficiently separated from their corresponding PI3K enzymatic products through either thin layer chromatography (TLC) or capillary electrophoresis (CE), and can be detected with high sensitivity by fluorescence. The biophysical and kinetic properties of the two probes are measured, and their suitability to characterize PI3K inhibitors is explored. Both probes show similar capacity as PI3K substrates for inhibitor characterization, yet also possess distinct properties that may suggest their different applications. These characterizations have laid the groundwork to systematically measure cellular PI3K activity, and have the potential to generate molecular fingerprints for diagnostic and therapeutic applications.
Authors:
Weigang Huang; Dechen Jiang; Xiaoyang Wang; Kelong Wang; Christopher E Sims; Nancy L Allbritton; Qisheng Zhang
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural     Date:  2011-07-26
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  401     ISSN:  1618-2650     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2011 Oct 
Date Detail:
Created Date:  2011-09-14     Completed Date:  2012-01-06     Revised Date:  2014-09-13    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1881-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Chromatography, Thin Layer / methods
Electrophoresis, Capillary / methods
Enzyme Inhibitors / pharmacology
Fluorescent Dyes / chemistry,  metabolism*
Kinetics
Phosphatidylinositol 3-Kinase / antagonists & inhibitors,  metabolism*
Phosphatidylinositol 4,5-Diphosphate / chemistry,  metabolism*
Sensitivity and Specificity
Grant Support
ID/Acronym/Agency:
CA139599/CA/NCI NIH HHS; R01 CA139599/CA/NCI NIH HHS; R01 CA139599-03/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Fluorescent Dyes; 0/Phosphatidylinositol 4,5-Diphosphate; EC 2.7.1.137/Phosphatidylinositol 3-Kinase
Comments/Corrections

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