Differential sensitivity of types 1 and 2 cholecystokinin receptors to membrane cholesterol

Ross M. Potter, Kaleeckal G. Harikumar, S. Vincent Wu, Laurence J Miller

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Recent studies indicate that membrane cholesterol can associate with G protein-coupled receptors (GPCRs) and affect their function. Previously, we reported that manipulation of membrane cholesterol affects ligand binding and signal transduction of the type 1 cholecystokinin receptor (CCK1R), a Class A GPCR. We now demonstrate that the closely related type 2 cholecystokinin receptor (CCK2R) does not share this cholesterol sensitivity. The sequences of both receptors reveal almost identical cholesterol interaction motifs in analogous locations in transmembrane segments two, three, four, and five. The disparity in cholesterol sensitivity between these receptors, despite their close structural relationship, provides a unique opportunity to define the possible structural basis of cholesterol sensitivity of CCK1R. To evaluate the relative contributions of different regions of CCK1R to cholesterol sensitivity, we performed ligand binding studies and biological activity assays of wildtype and CCK2R/CCK1R chimeric receptor-bearing Chinese hamster ovary cells after manipulation of membrane cholesterol. We also extended these studies to site-directed mutations within the cholesterol interaction motifs. The results contribute to a better understanding of the structural requirements for cholesterol sensitivity in CCK1R and provides insight into the function of other cholesterol-sensitive Class A GPCRs.

Original languageEnglish (US)
Pages (from-to)137-148
Number of pages12
JournalJournal of Lipid Research
Volume53
Issue number1
DOIs
StatePublished - Jan 2012

Fingerprint

Cholecystokinin B Receptor
Cholesterol
Membranes
G-Protein-Coupled Receptors
Bearings (structural)
Cholecystokinin Receptors
Ligands
Signal transduction
Cricetulus
Bioactivity
Biological Assay
Ovary
Assays
Signal Transduction

Keywords

  • Biological activity
  • Chimeric receptors
  • G protein-coupled receptors
  • Perfringolysin
  • Receptor binding

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

Differential sensitivity of types 1 and 2 cholecystokinin receptors to membrane cholesterol. / Potter, Ross M.; Harikumar, Kaleeckal G.; Wu, S. Vincent; Miller, Laurence J.

In: Journal of Lipid Research, Vol. 53, No. 1, 01.2012, p. 137-148.

Research output: Contribution to journalArticle

Potter, Ross M. ; Harikumar, Kaleeckal G. ; Wu, S. Vincent ; Miller, Laurence J. / Differential sensitivity of types 1 and 2 cholecystokinin receptors to membrane cholesterol. In: Journal of Lipid Research. 2012 ; Vol. 53, No. 1. pp. 137-148.
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