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Abstract
Grant Number: 1R43GM067365-01 PI Name: FENG, LI PI Email: lfeng@echelon-inc.com PI Title: Project Title: Fluorogenic Assays for Phospholipid Detection Abstract: DESCRIPTION (provided by applicant): Phospholipids (PL) including phosphoinositides (PIPn) are crucial structural and signaling molecules for proper inter- and intra-cellular communication in an organism. The disruption of lipid homeostasis is associated with numerous human pathologies. High levels of LPA, LPC, PI (3,4,5) P3, PI (3) P and PI (4,5) P2 are linked to specific pathologies such as cancer, myotubular myopathy and Lowe's syndrome. The inter-conversions of phospholipids are mediated by the interplay of phospholipases, kinases, phosphatases and other lipid binding proteins. It is important to analyze and quantify cellular and extra-cellular phospholipids to measure enzyme activities associated with specific pathologies. We propose to develop a fluorogenic assay based on chemically or genetically modified proteins that selectively recognize PLs and PIPns. The modification will be site-specific and will incorporate novel light sensitive entities. These reagents will be applied in Phase II to develop high throughput screening methods for rapid and automated assay of lipid kinases and phosphatases. Such reagents can also be used in the development of cell-based assays.
Thesaurus Terms:
chemical conjugate, fluorescence, fluorescent dye /probe, phosphatidylinositol, phospholipid, protein engineering, recombinant protein, technology /technique development
glutathione transferase, protein purification, second messenger
Institution: ECHELON BIOSCIENCES, INC. 675 ARAPEEN DR, STE 302 SALT LAKE CITY, UT 84108 Fiscal Year: 2003 Department: Project Start: 01-FEB-2003 Project End: 31-JUL-2003 ICD: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES IRG: ZRG1
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