Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 291, Issue 42, Pages 21925-21944Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M116.751362
Keywords
G protein-coupled receptor (GPCR); molecular dynamics; molecular modeling; signal transduction; yeast; adrenomedullin; adrenomedullin 2; CGRP; RAMP; signal bias
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Funding
- National Heart Foundation of New Zealand
- School of Biological Sciences, University of Auckland [BB/M00015X/1, BB/M000176/1, BB/M006883/1]
- BBSRC
- BBSRC Doctoral Training Partnership Grant [BB/JO14540/1]
- MRC Doctoral Training Partnership [MR/J003964/1]
- Warwick Impact Fund
- Warwick Research Development Fund [RD13301]
- Warwick Undergraduate Research Scholarship Scheme
- Biotechnology and Biological Sciences Research Council [1643678, BB/M007529/1, BB/M000176/1, BB/M00015X/2, BB/M006883/1, BB/M00015X/1] Funding Source: researchfish
- Medical Research Council [1361423] Funding Source: researchfish
- BBSRC [BB/M007529/1, BB/M00015X/2, BB/M00015X/1, 1643678, BB/M000176/1, BB/M006883/1] Funding Source: UKRI
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The calcitonin gene-related peptide (CGRP) family of G protein-coupled receptors (GPCRs) is formed through the association of the calcitonin receptor-like receptor (CLR) and one of three receptor activity-modifying proteins (RAMPs). Binding of one of the three peptide ligands, CGRP, adrenomedullin (AM), and intermedin/adrenomedullin 2 (AM2), is well known to result in a G(s)-mediated increase in cAMP. Here we used modified yeast strains that couple receptor activation to cell growth, via chimeric yeast/G subunits, and HEK-293 cells to characterize the effect of different RAMP and ligand combinations on this pathway. We not only demonstrate functional couplings to both G(s) and G(q) but also identify a G(i) component to CLR signaling in both yeast and HEK-293 cells, which is absent in HEK-293S cells. We show that the CGRP family of receptors displays both ligand- and RAMP-dependent signaling bias among the G(s), G(i), and G(q/11) pathways. The results are discussed in the context of RAMP interactions probed through molecular modeling and molecular dynamics simulations of the RAMP-GPCR-G protein complexes. This study further highlights the importance of RAMPs to CLR pharmacology and to bias in general, as well as identifying the importance of choosing an appropriate model system for the study of GPCR pharmacology.
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