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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Gibson, Scott K. Gilman, Alfred G. |
| Description | Author Affiliation: Gibson SK ( Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. scott.gibson@utsouthwestern.edu); |
| Abstract | Previous studies of the specificity of receptor interactions with G protein subunits in living cells have relied on measurements of second messengers or other downstream responses. We have examined the selectivity of interactions between alpha2-adrenergic receptors (alpha2R) and various combinations of Gialpha and Gbeta subunit isoforms by measuring changes in FRET between Gialpha-yellow fluorescent protein and cyan fluorescent protein-Gbeta chimeras in HeLa cells. All combinations of Gialpha1, -2, or -3 with Gbeta1, -2, or -4 were activated to some degree by endogenous alpha2Rs as judged by agonist-dependent decreases in FRET. The degree of G protein activation is determined by the combination of Gialpha and Gbeta subunits rather than by the identity of an individual subunit. RT-PCR analysis and small interfering RNA knockdown of alpha2R subtypes, followed by quantification of radiolabeled antagonist binding, demonstrated that HeLa cells express alpha2a- and alpha2b-adrenergic receptor isoforms in a 2:1 ratio. Increasing receptor number by overexpression of the alpha2aR subtype minimized the differences among coupling preferences for Gialpha and Gbeta isoforms. The molecular properties of each Gialpha, Gbeta, and alpha2-adrenergic receptor subtype influence signaling efficiency for the alpha2-adrenergic receptor-mediated signaling pathway. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 1 |
| Volume Number | 103 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2006-01-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | GTP-Binding Protein Alpha Subunits, Gi-Go Metabolism GTP-Binding Protein Beta Subunits Receptors, Adrenergic, Alpha-2 Signal Transduction Physiology Enzyme Activation Fluorescence Resonance Energy Transfer Fluorescent Dyes HeLa Cells Microscopy, Fluorescence RNA, Small Interfering Genetics Reverse Transcriptase Polymerase Chain Reaction Substrate Specificity Comparative Study Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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