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Protein Overlays for Phosphoinositide Detection

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: N/A
Agency Tracking Number: 1R43GM064251-01
Amount: $100,000.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: N/A
Solicitation Number: N/A
Timeline
Solicitation Year: N/A
Award Year: 2001
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
420 CHIPETA WAY, STE 180
SALT LAKE CITY, UT 84108
United States
DUNS: N/A
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 COLIN FERGUSON
 () -
Business Contact
Phone: (801) 588-0455
Email: GTHEATH@ECHELON-INC.COM
Research Institution
N/A
Abstract

DESCRIPTION (provided by applicant): Phosphoinositides (PIPns), e.g.,
phosphatidylinositol 4,5-bisphosphate (PI 4,5) P2) are key signaling molecules
in cellular communication. Specific PIPns recruit target signaling proteins and
intracellular adapter proteins to inner membrane sites to activate protein
kinases and thereby initiate signal transduction cascades in order to regulate
exo- and endocytosis, protein sorting, cell proliferation and migration,
tumorigenesis, apoptosis, and the control of cell shape. Determining the
selectivity of a given target protein for a specific PIPn has become an
important issue in deciphering downstream effectors and in new drug discovery
efforts for potential therapeutics that could control protein-PIPn
interactions.

We propose to develop both components of an efficient protein overlay system
that allows determining lipid selectivity of a known protein or levels of a
given PIPn produced by lipid kinase and phosphatase reactions. Both methods use
the same principle, i.e., detection of proteins bound to PIPns immobilized on
nitrocellulose. First, we will optimize the lipid blot method, which we call
"PIP-Strips (TM)." Second, we will develop highly selective, high-affinity
proteins that recognize a specific PIPn using two coupled pleckstrin homology
(PH) or FYVE domains; this new strategy is called Double PH(tm). The reagents
prepared and optimized in this Phase I project will be used in Phase II to
develop a variety of high throughput screening methods, e.g., rapid, automated
assays for PIPn kinases and phosphatase and screening for small molecule
inhibitors of protein-PIPn binding
PROPOSED COMMERCIAL APPLICATION:
A number of commercial applications have been envisioned for the products developed
in this grant. PIP-Strips(TM) will be primarily sold as tools to determine lipid binding
specificity of isolated proteins, their original purpose. Other applications include antibody
screening and in house quality control of cloned proteins. Lipo-PIP-Strips(TM) will also fill
these same roles. The Double-PH reagents will be sold as individual items targeted to a
specific lipid and a kits focussing on a class of lipids such as 3'-phosphorylated PIPns.

* Information listed above is at the time of submission. *

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