Structure-function studies of the eighth hydrophobic domain of a serotonin receptor

Joyce Hurley, Laura J. Bloem, Fredrick Pavalko, Jian Liu, Mingting Tian, Jay R. Simon, Lei Yu

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The most prominent structural feature of the G protein-coupled receptor superfamily is their seven hydrophobic domains, which are postulated to form membrane-spanning α helices. Some members of the G protein-coupled receptor family, specifically several serotonin (5-HT) receptors, possess eight hydrophobic domains. The importance of this extra hydrophobic domain, located at the N terminus of the receptor, is unknown. This question was addressed by deleting the extrahydrophobic region from the 5-HT(2C) receptor and comparing its function and topology with those of the wild-type receptor. Immunofluorescence microscopy was used to determine the location of the N terminus of the epitope-tagged wild-type and mutant receptors. The N terminus of both receptors was extracellular, suggesting that the extra hydrophobic domain does not change the topology of this receptor and is unlikely to be a membrane-spanning α helix. Radioligand-binding studies in transfected cells and expression studies in Xenopus oocytes demonstrated that seven hydrophobic domains were sufficient for normal function in these assays. Interestingly, the mutant receptor, now containing seven hydrophobic domains, is expressed at higher levels in transfected cells than the wild-type receptor containing eight hydrophobic domains, suggesting that the extra hydrophobic domain does impact the activity of this receptor by regulating its expression.

Original languageEnglish
Pages (from-to)413-421
Number of pages9
JournalJournal of Neurochemistry
Volume72
Issue number1
DOIs
StatePublished - 1999

Fingerprint

Serotonin Receptors
G-Protein-Coupled Receptors
Topology
Receptor, Serotonin, 5-HT2C
Membranes
Xenopus
Fluorescence Microscopy
Oocytes
Epitopes
Assays
Serotonin
Microscopic examination
serotonin 5 receptor

Keywords

  • α helix
  • G protein-coupled receptor
  • Hydrophobic domain
  • Topology

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Structure-function studies of the eighth hydrophobic domain of a serotonin receptor. / Hurley, Joyce; Bloem, Laura J.; Pavalko, Fredrick; Liu, Jian; Tian, Mingting; Simon, Jay R.; Yu, Lei.

In: Journal of Neurochemistry, Vol. 72, No. 1, 1999, p. 413-421.

Research output: Contribution to journalArticle

Hurley, Joyce ; Bloem, Laura J. ; Pavalko, Fredrick ; Liu, Jian ; Tian, Mingting ; Simon, Jay R. ; Yu, Lei. / Structure-function studies of the eighth hydrophobic domain of a serotonin receptor. In: Journal of Neurochemistry. 1999 ; Vol. 72, No. 1. pp. 413-421.
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