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

Conner, Matthew; Hicks, Matthew R.; Dafforn, Tim; Knowles, Timothy J.; Ludwig, Christian; Staddon, Susan; Overduin, Michael; Günther, Ulrich L.; Thome, Johannes; Wheatley, M.; Poyner, David R. and Conner, Alex C. (2008). Functional and biophysical analysis of the C-terminus of the CGRP-receptor; a family B GPCR. Biochemistry, 47 (32), pp. 8434-8444.

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 DOI: https://doi.org/10.1021/bi8004126
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Life & Health Sciences > Applied Health Research Group
Life & Health Sciences > Biomedical Sciences research group
Life & Health Sciences > Pharmacy
Related URLs:
Uncontrolled Keywords: G-protein coupled receptors,GPCRs,C-terminus,membrane-parallel,eighth helix,mutations,N400,C436,CGRP-mediated internalization,G388,W399),receptor localization,cell surface,dichroism,waterLOGSY,NMR spectroscopy,SALMON,peptide,interfacial tryptophan residue,Biochemistry
Published Date: 2008-07-18

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