Effect of multiple serine/alanine mutations in the transmembrane spanning region V of the D-2 dopamine receptor on ligand binding

Coley, Clare and Woodward, Robert and Johansson, Anette M. and Strange, Philip G. and Naylor, Louise H. (2000) Effect of multiple serine/alanine mutations in the transmembrane spanning region V of the D-2 dopamine receptor on ligand binding. Journal of Neurochemistry, 74 (1). pp. 358-366. ISSN 0022-3042. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1046/j.1471-4159.2000.0740358...

Abstract

Three conserved serine residues (Ser(193), Ser(194), and Ser(197)) in transmembrane spanning region (TM) V of the D-2 dopamine receptor have been mutated to alanine, individually and in combination, to explore their role in ligand binding and G protein coupling. The multiple Ser --> Ala mutations had no effect on the binding of most antagonists tested, including [H-3]spiperone, suggesting that the multiple mutations did not affect the overall conformation of the receptor protein. Double or triple mutants containing an Ala(197) mutation showed a decrease in affinity for domperidone, whereas Ala(193) mutants showed an increased affinity for a substituted benzamide, remoxipride, However, dopamine showed large decreases in affinity (>20-fold) for each multiple mutant receptor containing the Ser(193)Ala mutation, and the high-affinity (coupled) state of the receptor (in the absence of GTP) could not be detected for any of the multiple mutants. A series of monohydroxylated phenylethylamines and aminotetralins was tested for their binding to the native and multiple mutant D-2 dopamine receptors. The results obtained suggest that Ser(193) interacts with the hydroxyl of S-5-hydroxy-2-dipropylaminotetralin (OH-DPAT) and Ser(197) with the hydroxyl of R-5-OH-DPAT. We predict that Ser(193) interacts with the hydroxyl of R-7-OH-DPAT and the 3-hydroxyl (m-hydroxyl) of dopamine. Therefore, the conserved serine residues in TMV of the D-2 dopamine receptor are involved in hydrogen bonding interactions with selected antagonists and most agonists tested and also enable agonists to stabilise receptor-G protein coupling.

Item Type: Article
Uncontrolled keywords: D-2 dopamine receptor; ligand binding; serine residues; mutagenesis
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: P. Ogbuji
Date Deposited: 17 Apr 2009 18:34
Last Modified: 16 Apr 2014 15:17
Resource URI: https://kar.kent.ac.uk/id/eprint/16299 (The current URI for this page, for reference purposes)
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