Members of the large biogenic amine family of GPCRs are involved in several medically relevant physiological processes and, as such, are major therapeutic targets. Current pharmacological and transgenic techniques are unsuitable for detailed functional studies of a single receptor in a specific cell or tissue in vivo. The focus of this proposal is to develop novel receptor mutants which are selectively activated by a biologically-inert ligand. A """"""""designer"""""""" receptor used in combination with traditional pharmacology and transgenics approaches will reveal novel intra- and intercellular functions for the receptor. To this end, a yeast-based screen will be employed to identify acetylcholine muscarinic 3 receptor mutants that selectively respond to a foreign drug versus its native ligand. Upon initial characterization of drug potency and efficacy in yeast, the mutant receptor will be expressed in human cells to verify that activation of the designer receptor by the designer drug can initiate indigenous physiological responses, including PIP2 hydrolysis, ERK-1/2 phosphorylation and smooth muscle cell contraction. Finally, bioinformatics, including multi-receptor molecular modeling, will be used to rationally engineer pan-bioamine family designer receptors.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM074554-02
Application #
7062444
Study Section
Special Emphasis Panel (ZRG1-F03B (20))
Program Officer
Fabian, Miles
Project Start
2005-04-01
Project End
2006-05-31
Budget Start
2006-04-01
Budget End
2006-05-31
Support Year
2
Fiscal Year
2006
Total Cost
$13,605
Indirect Cost
Name
Case Western Reserve University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
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Armbruster, Blaine N; Li, Xiang; Pausch, Mark H et al. (2007) Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand. Proc Natl Acad Sci U S A 104:5163-8
Keiser, Michael J; Roth, Bryan L; Armbruster, Blaine N et al. (2007) Relating protein pharmacology by ligand chemistry. Nat Biotechnol 25:197-206