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METHOD DEVELOPMENT AND INVESTIGATION OF FLUORESCENT PHOSPHOINOSITIDE CELL SIGNALING PROPERTIES BY CAPILLARY ELECTROPHORESIS

Quainoo, Emmanuel W0bil
http://rave.ohiolink.edu/etdc/view?acc_num=kent1271693846

Abstract Details

2010, MS, Kent State University, College of Arts and Sciences / Department of Chemistry.
Phosphatidyl inositides (PIs) are important regulators of cell signaling. Phosphoinositide 3-kinase (PI3-K)-activated signaling plays a key role in the development of cancer. Therefore various anticancer treatments target this pathway. The aim of this work was to develop an optimized method to separate fluorescent PIPs quickly and efficiently at room temperature using CE-LIF. The reason for the development of a method capable of separating the PIPs at room temperature was that, it could be used for single cell studies. In this regard, the effect of various cations on the separation of the PIs and their seven PIP derivatives was investigated. The effect of pH, temperature, voltage, as well as other buffer mixtures were also studied. Based on this developed method, the activity of PI3-K and PTEN enzymes in NIH 3T3 cells and MDA-MB 231 cells was also studied. The bioactivity of PIPs in these cells was investigated to ascertain whether preliminary CE studies would prove that they were able to be converted inside the cells into other phosphorylated derivatives. Lipid kinases and phosphatases play active roles in cell signaling. These have serious implications in many disease states. For example, accumulation of PIP3 leads to metastatic cancers. Therefore, inhibition studies were also performed to find out whether PI3-K inhibitors were able to block the conversion of PIP2 to PIP3. This is of particular interest in cell-based assays and research involving cancer drug development.
Simon M Mwongela, PhD (Advisor)
Songping Huang, PhD (Committee Member)
Bansidhar Datta, PhD (Committee Member)
90 p.
Biochemistry
phosphoinositides; capillary electrophoresis; phosphatidyl inositol; phosphatidyl inositol phosphates; micellar electrokinetic chromatography; metal cations; inhibitors; phosphatases; cell signaling; lipids; PTEN; PI3-K; PIPS

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Citations

  • Quainoo, E. W. (2010). METHOD DEVELOPMENT AND INVESTIGATION OF FLUORESCENT PHOSPHOINOSITIDE CELL SIGNALING PROPERTIES BY CAPILLARY ELECTROPHORESIS [Master's thesis, Kent State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=kent1271693846

    APA Style (7th edition)

  • Quainoo, Emmanuel. METHOD DEVELOPMENT AND INVESTIGATION OF FLUORESCENT PHOSPHOINOSITIDE CELL SIGNALING PROPERTIES BY CAPILLARY ELECTROPHORESIS. 2010. Kent State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=kent1271693846.

    MLA Style (8th edition)

  • Quainoo, Emmanuel. "METHOD DEVELOPMENT AND INVESTIGATION OF FLUORESCENT PHOSPHOINOSITIDE CELL SIGNALING PROPERTIES BY CAPILLARY ELECTROPHORESIS." Master's thesis, Kent State University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=kent1271693846

    Chicago Manual of Style (17th edition)

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This open access ETD is published by Kent State University and OhioLINK.