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PMID:7706278
| Citation |
Guiramand, J, Montmayeur, JP, Ceraline, J, Bhatia, M and Borrelli, E (1995) Alternative splicing of the dopamine D2 receptor directs specificity of coupling to G-proteins. J. Biol. Chem. 270:7354-8 |
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| Abstract |
Two isoforms of the dopamine D2 receptor have been characterized, D2L (long) and D2S (short), generated by alternative splicing from the same gene. They differ by an in-frame insert of 29 amino acids specific to D2L within the putative third intracytoplasmic loop of the receptor. We have previously demonstrated (Montmayeur, J.-P., Guiramand, J., and Borelli, E. (1993) Mol. Endocrinol. 7, 161-170) that D2S and D2L, although presenting very similar pharmacological profiles, couple differently to the alpha-subunit of guanine nucleotide-binding regulatory proteins (G-proteins). In particular, D2L, but not D2S, requires the presence of the alpha-subunit of the inhibitory G-protein (G alpha i2) to elicit greater inhibition of adenylyl cyclase activity. The insert present in D2L must therefore confer the specificity of interaction with G alpha i2. Thus, we introduced substitution mutations within the D2L insert. These mutant receptors were expressed in JEG3 cells, a G alpha i2-deficient cell line, scoring for those presenting an increased inhibition of adenylyl cyclase by dopamine. Our analysis identified two mutants, S259/262A and D249V, with these properties. These results clearly show that the insert present in D2L plays a critical role in the selectivity for the G-proteins interacting with the receptor. |
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| Keywords |
Adenylate Cyclase/antagonists & inhibitors; Alternative Splicing; Amino Acid Sequence; Animals; Apomorphine/analogs & derivatives; Apomorphine/metabolism; Base Sequence; Cell Line; Cloning, Molecular; Cyclic AMP/metabolism; Dopamine/pharmacology; Dopamine Agonists/metabolism; GTP-Binding Proteins/metabolism; Humans; Isoproterenol/metabolism; Kinetics; Mice; Molecular Sequence Data; Mutagenesis, Insertional; Receptors, Adrenergic, beta-2/biosynthesis; Receptors, Adrenergic, beta-2/metabolism; Receptors, Dopamine D2/biosynthesis; Receptors, Dopamine D2/metabolism; Recombinant Proteins/biosynthesis; Recombinant Proteins/metabolism; Spiperone/metabolism; Transfection |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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