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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Jin, Xiaochun Steinbach, Joe Henry Blanton, Michael P. Hamouda, Ayman K. Srivastava, Shouryadeep Pandhare, Akash Sanghvi, Mitesh Duddempudi, Phaneendra K. |
| Description | Author Affiliation: Hamouda AK ( Department of Pharmacology and Neuroscience and the Center for Membrane Protein Research, School of Medicine, Texas Tech University Health Sciences Center, 3601 4th Street, Lubbock, TX 79430, USA.) |
| Abstract | The development of nicotinic acetylcholine receptor (nAChR) agonists, particularly those that discriminate between neuronal nAChR subtypes, holds promise as potential therapeutic agents for many neurological diseases and disorders. To this end, we photoaffinity labeled human alpha4beta2 and rat alpha4beta4 nAChRs affinity-purified from stably transfected HEK-293 cells, with the agonists [(125)I]epibatidine and 5[(125)I]A-85380. Our results show that both agonists photoincorporated into the beta4 subunit with little or no labeling of the beta2 and alpha4 subunits respectively. [(125)I]epibatidine labeling in the beta4 subunit was mapped to two overlapping proteolytic fragments that begin at beta4V102 and contain Loop E (beta4I109-P120) of the agonist binding site. We were unable to identify labeled amino acid(s) in Loop E by protein sequencing, but we were able to demonstrate that beta4Q117 in Loop E is the principal site of [(125)I]epibatidine labeling. This was accomplished by substituting residues in the beta2 subunit with the beta4 homologs and finding [(125)I]epibatidine labeling in beta4 and beta2F119Q subunits with little, if any, labeling in alpha4, beta2, or beta2S113R subunits. Finally, functional studies established that the beta2F119/beta4Q117 position is an important determinant of the receptor subtype-selectivity of the agonist 5I-A-85380, affecting both binding affinity and channel activation. |
| ISSN | 00063002 |
| Journal | Biochimica et Biophysica Acta (BBA) - Reviews on Cancer |
| Issue Number | 9 |
| Volume Number | 1788 |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2009-09-01 |
| Publisher Place | Netherlands |
| Access Restriction | Open |
| Subject Keyword | Azetidines Metabolism Bicyclo Compounds, Heterocyclic Nicotinic Agonists Pyridines Receptors, Nicotinic Chemistry Amino Acid Sequence Animals Binding Sites Cells, Cultured Iodine Radioisotopes Molecular Sequence Data Pharmacology Oocytes Physiology Photoaffinity Labels Drug Effects Genetics Sequence Alignment Xenopus Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Biochemistry |
| Content Type | Text |
| Resource Type | Article |
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