Functional and biophysical analysis of the C-terminus of the CGRP-receptor; a family B GPCR

Biochemistry. 2008 Aug 12;47(32):8434-44. doi: 10.1021/bi8004126. Epub 2008 Jul 18.

Abstract

G-protein coupled receptors (GPCRs) typically have a functionally important C-terminus which, in the largest subfamily (family A), includes a membrane-parallel eighth helix. Mutations of this region are associated with several diseases. There are few C-terminal studies on the family B GPCRs and no data supporting the existence of a similar eighth helix in this second major subfamily, which has little or no sequence homology to family A GPCRs. Here we show that the C-terminus of a family B GPCR (CLR) has a disparate region from N400 to C436 required for CGRP-mediated internalization, and a proximal region of twelve residues (from G388 to W399), in a similar position to the family A eighth helix, required for receptor localization at the cell surface. A combination of circular and linear dichroism, fluorescence and modified waterLOGSY NMR spectroscopy (SALMON) demonstrated that a peptide mimetic of this domain readily forms a membrane-parallel helix anchored to the liposome by an interfacial tryptophan residue. The study reveals two key functions held within the C-terminus of a family B GPCR and presents support for an eighth helical region with striking topological similarity to the nonhomologous family A receptor. This helix structure appears to be found in most other family B GPCRs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Calcitonin Gene-Related Peptide / chemistry
  • Calcitonin Gene-Related Peptide / metabolism*
  • Chlorocebus aethiops
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Receptors, Calcitonin Gene-Related Peptide / biosynthesis
  • Receptors, Calcitonin Gene-Related Peptide / chemistry*
  • Receptors, Calcitonin Gene-Related Peptide / genetics
  • Receptors, Calcitonin Gene-Related Peptide / physiology*

Substances

  • Receptors, Calcitonin Gene-Related Peptide
  • Calcitonin Gene-Related Peptide