Identification of N-terminal receptor activity-modifying protein residues important for calcitonin gene-related peptide, adrenomedullin, and amylin receptor function

Show simple item record Qi, T en Christopoulos, G en Bailey, Richard en Christopoulos, A en Sexton, PM en Hay, Deborah en
dc.coverage.spatial United States en 2012-03-30T01:50:19Z en 2008-10 en
dc.identifier.citation Molecular Pharmacology 74(4):1059-1071 2008 en
dc.identifier.issn 0026-895X en
dc.identifier.uri en
dc.description.abstract Calcitonin-family receptors comprise calcitonin receptor-like receptor (CL) or calcitonin receptor and receptor activity-modifying protein (RAMP) pairings. Calcitonin gene-related peptide (CGRP) receptors are CL/RAMP1, whereas adrenomedullin (AM) receptors are CL/RAMP2 (AM1 receptor) or CL/RAMP3 (AM2 receptor). Amylin (Amy) receptors are RAMP hetero-oligomers with the calcitonin receptor (AMY1, AMY2, and AMY3, respectively). How RAMPs change G protein-coupled receptor pharmacology is not fully understood. We exploited sequence differences between RAMP1 and RAMP3 to identify individual residues capable of altering receptor pharmacology. Alignment of human RAMPs revealed eight residues that are conserved in RAMP2 and RAMP3 but are different in RAMP1. We hypothesized that residues in RAMP2 and RAMP3, but not RAMP1, are responsible for making CL/RAMP2 and CL/RAMP3 AM receptors. Using site-directed mutagenesis, we introduced individual RAMP3 residues into RAMP1 and vice versa in these eight positions. Mutant or wild-type RAMPs were transfected into Cos7 cells with CL or the insert-negative form of the calcitonin receptor [CT(a)]. Agonist-stimulated cAMP production and cell-surface expression of constructs were measured. Position 74 in RAMP1 and RAMP3 was critical for determining AM potency and affinity, and Phe93 in RAMP1 was an important contributor to alphaCGRP potency at CGRP receptors. Mutant RAMP/CT(a) receptor complexes displayed different phenotypes. It is noteworthy that RAMP1 S103N and W74E mutations led to enhanced rAmy potency, probably related to increased cell-surface expression of these complexes. This differs from the effect on CL-based receptors where expression was unchanged. Targeted substitution has emphasized the importance of position 74 in RAMP1/RAMP3 as a key determinant of AM pharmacology. en
dc.language eng en
dc.publisher The American Society for Pharmacology and Experimental Therapeutic en
dc.relation.ispartofseries Molecular Pharmacology en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from en
dc.rights.uri en
dc.subject Adrenomedullin en
dc.subject Amino Acid Sequence en
dc.subject Amino Acid Substitution en
dc.subject Animals en
dc.subject COS Cells en
dc.subject Calcitonin Gene-Related Peptide en
dc.subject Cells, Cultured en
dc.subject Cercopithecus aethiops en
dc.subject Conserved Sequence en
dc.subject Cyclic AMP en
dc.subject Forskolin en
dc.subject Hemagglutinins en
dc.subject Humans en
dc.subject Immunoglobulin G en
dc.subject Inhibitory Concentration 50 en
dc.subject Intracellular Signaling Peptides and Proteins en
dc.subject Membrane Proteins en
dc.subject Molecular Sequence Data en
dc.subject Protein Sorting Signals en
dc.subject Receptor Activity-Modifying Protein 1 en
dc.subject Receptor Activity-Modifying Protein 2 en
dc.subject Receptor Activity-Modifying Protein 3 en
dc.subject Receptor Activity-Modifying Proteins en
dc.subject Receptors, Islet Amyloid Polypeptide en
dc.subject Receptors, Peptide en
dc.subject Sequence Homology, Amino Acid en
dc.subject Transfection en
dc.title Identification of N-terminal receptor activity-modifying protein residues important for calcitonin gene-related peptide, adrenomedullin, and amylin receptor function en
dc.type Journal Article en
dc.identifier.doi 10.1124/mol.108.047142 en
pubs.issue 4 en
pubs.begin-page 1059 en
pubs.volume 74 en
dc.rights.holder Copyright: The American Society for Pharmacology and Experimental Therapeutic en
dc.identifier.pmid 18593822 en
pubs.end-page 1071 en
dc.rights.accessrights en
pubs.subtype Article en
pubs.elements-id 115977 en Science en Biological Sciences en Science Research en Maurice Wilkins Centre (2010-2014) en
dc.identifier.eissn 1521-0111 en
dc.identifier.pii mol.108.047142 en
pubs.record-created-at-source-date 2012-02-21 en
pubs.dimensions-id 18593822 en

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