Nicotinic acetylcholine receptors (AChRs) are multimeric proteins assembled from a wide variety of subunits. Each combination of subunits has unique pharmacological and physiological properties, resulting in many different receptor subtypes that could serve different functional roles. Agonists such as acetylcholine activate the channel by binding to at least two subunits in the receptor. All of the different subunits have a unique disulfide bond near the ACh binding site, and when this disulfide is reduced, nicotinic receptor function is largely blocked. The applicant proposes to use a novel set of labeling and cross-linking agents based on arsenical chemistry in order to analyze the subunit composition of receptors using subunit-specific antibodies. These agents are designed to potently and covalently attach to the reduced disulfide. The applicant proposes to make radioiodinated and biotinylated arsenical ligands, show that they label the Torpedo AChR (as a model), and then test them on neuronal receptors in the chick nervous system. In addition, a series of photoactivatable crosslinking ligands will also be used to determine the subunits adjacent to the subunits. The applicant will also use a series of biotinylated arsenical reagents to map the distance from the disulfide bond in the binding site from the surface of the receptor.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS022472-13
Application #
2635684
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Program Officer
Kitt, Cheryl A
Project Start
1985-09-01
Project End
1999-11-30
Budget Start
1998-01-01
Budget End
1998-12-31
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Northeastern University
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02115