The membrane proximal region of the cannabinoid receptor CB1 N-terminus can allosterically modulate ligand affinity

Jonathan F. Fay, David Farrens

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The human cannabinoid receptor, CB1, a G protein-coupled receptor (GPCR), contains a relatively long (∼110 a.a.) amino terminus, whose function is still not defined. Here we explore a potential role for the CB1 N-terminus in modulating ligand binding to the receptor. Although most of the CB1 N-terminus is not necessary for ligand binding, previous studies have found that mutations introduced into its conserved membrane proximal region (MPR) do impair the receptors ability to bind ligand. Moreover, within the highly conserved MPR (∼ residues 90-110) lie two cysteine residues that are invariant in all CB1 receptors. We find these two cysteines (C98 and C107) form a disulfide in heterologously expressed human CB1, and this C98-C107 disulfide is much more accessible to reducing agents than the previously known disulfide in extracellular loop 2 (EL2). Interestingly, the presence of the C 98-C107 disulfide modulates ligand binding to the receptor in a way that can be quantitatively analyzed by an allosteric model. The C 98-C107 disulfide also alters the effects of allosteric ligands for CB1, Org 27569 and PSNCBAM-1. Together, these results provide new insights into how the N-terminal MPR and EL2 act together to influence the high-affinity orthosteric ligand binding site in CB1 and suggest that the CB1 N-terminal MPR may be an area through which allosteric modulators can act.

Original languageEnglish (US)
Pages (from-to)8286-8294
Number of pages9
JournalBiochemistry
Volume52
Issue number46
DOIs
StatePublished - Nov 19 2013

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Cannabinoid Receptor CB1
Disulfides
Ligands
Membranes
Cysteine
Reducing Agents
G-Protein-Coupled Receptors
Modulators
Binding Sites
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

The membrane proximal region of the cannabinoid receptor CB1 N-terminus can allosterically modulate ligand affinity. / Fay, Jonathan F.; Farrens, David.

In: Biochemistry, Vol. 52, No. 46, 19.11.2013, p. 8286-8294.

Research output: Contribution to journalArticle

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