The effect of mutations in the DRY motif on the constitutive activity and structural instability of the histamine H(2) receptor.

Détails

ID Serval
serval:BIB_D280983F5375
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
The effect of mutations in the DRY motif on the constitutive activity and structural instability of the histamine H(2) receptor.
Périodique
Molecular Pharmacology
Auteur(s)
Alewijnse A.E., Timmerman H., Jacobs E.H., Smit M.J., Roovers E., Cotecchia S., Leurs R.
ISSN
0026-895X (Print)
ISSN-L
0026-895X
Statut éditorial
Publié
Date de publication
2000
Volume
57
Numéro
5
Pages
890-898
Langue
anglais
Résumé
In previous studies we showed that the wild-type histamine H(2) receptor stably expressed in Chinese hamster ovary cells is constitutively active. Because constitutive activity of the H(2) receptor is already found at low expression levels (300 fmol/mg protein) this receptor is a relatively unique member of the G-protein-coupled receptor (GPCR) family and a useful tool for studying GPCR activation. In this study the role of the highly conserved DRY motif in activation of the H(2) receptor was investigated. Mutation of the aspartate 115 residue in this motif resulted in H(2) receptors with high constitutive activity, increased agonist affinity, and increased signaling properties. In addition, the mutant receptors were shown to be highly structurally instable. Mutation of the arginine 116 residue in the DRY motif resulted also in a highly structurally instable receptor; expression of the receptor could only be detected after stabilization with either an agonist or inverse agonist. Moreover, the agonist affinity at the Arg-116 mutant receptors was increased, whereas the signal transduction properties of these receptors were decreased. We conclude that the Arg-116 mutant receptors can adopt an active conformation but have a decreased ability to couple to or activate the G(s)-protein. This study examines the pivotal role of the aspartate and arginine residues of the DRY motif in GPCR function. Disruption of receptor stabilizing constraints by mutation in the DRY motif leads to the formation of active GPCR conformations, but concomitantly to GPCR instability.
Mots-clé
Amino Acid Motifs, Amino Acid Sequence, Animals, Arginine/genetics, Arginine/metabolism, Cells, Cultured, Cricetinae, Humans, Ligands, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Conformation, Ranitidine/pharmacology, Receptors, Adrenergic, alpha-1/genetics, Receptors, Adrenergic, alpha-1/metabolism, Receptors, Histamine H2/chemistry, Receptors, Histamine H2/genetics, Signal Transduction
Pubmed
Web of science
Création de la notice
24/01/2008 11:05
Dernière modification de la notice
20/08/2019 15:52
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