Crystal structure of the human receptor activity‐modifying protein 1 extracellular domain

Kusano, Shoichi; Kukimoto-Niino, Mutsuko; Akasaka, Ryogo; Toyama, Motomichi; Terada, Takaho; Shirouzu, Mikako; Shindo, Takayuki; Yokoyama, Shigeyuki

doi:10.1110/ps.036012.108

Cited by 51 publications

(70 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It belongs to a family of three receptor activity-modifying proteins (RAMP1, -2, and -3) that affect the glycosylation, membrane trafficking and ligand recognition properties of the calcitonin receptor-like receptor, a family B receptor distantly related to the glucagon receptor (33). However, similar to the homologous RAMP1 expressed in Escherichia coli (34), the four extracellular cysteines of RAMP2 are probably involved in two conserved disulfide bridges and therefore not available to cysteine reagents. RAMP2 is therefore unlikely to be responsible for the effect of MTSET on glucagon recognition in HEK cells.…”

Section: Discussionmentioning

confidence: 99%

Mutational and Cysteine Scanning Analysis of the Glucagon Receptor N-terminal Domain

Prévost

Vertongen

Raussens

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

The glucagon receptor belongs to the B family of G-protein coupled receptors. Little structural information is available about this receptor and its association with glucagon. We used the substituted cysteine accessibility method and three-dimensional molecular modeling based on the gastrointestinal insulinotropic peptide and glucagon-like peptide 1 receptor structures to study the N-terminal domain of this receptor, a central element for ligand binding and specificity. with at least one free cysteine associated with the receptor prevented glucagon recognition and that [2-(trimethylammonium)ethyl]methanethiosulfonate treatment relieved this inhibition. The substituted cysteine accessibility method was also performed on 15 residues selected using the three-dimensional models. Several receptor mutants, despite a relatively high predicted cysteine accessibility, could not be labeled by specific reagents. The three-dimensional models show that these mutated residues are located on one face of the protein. This could be part of the interface between the receptor and the unidentified inhibitory protein, making these residues inaccessible to biotinylation compounds.The G-protein coupled receptors related to the secretin receptor (1), known as "family B" receptors, form a very small but very important subfamily of G protein-coupled receptors. This family includes, among others, the receptors for parathyroid hormone, glucagon, gastrointestinal insulinotropic peptide (GIP), 4 glucagon-like peptide 1 (GLP-1), corticotrophinrelated factor (CRF), or growth hormone-releasing hormone. Although the two receptor families probably evolved from a common ancestor, rhodopsin-related (family A) and family B receptors present few similarities (1). Except for two cysteine residues in the first and second extracellular loops, none of the signature amino acids defining family A G protein-coupled receptors are identified in family B receptors. It is therefore impossible to extrapolate the structural information gathered on rhodopsin activation (2, 3) to these receptors.All family B receptors possess a large extracellular N-terminal domain of 100 -150 amino acids, which contains highly conserved residues and three conserved disulfide bonds. Truncated and hybrid peptide recognition by chimeric receptors indicate that these domains anchor the cognate agonist peptides through their C-terminal region, whereas the N-terminal regions of the peptides interact with and activate the receptors via the heptahelical transmembrane domains (4, 5).The three-dimensional structures of the N-terminal domain of the CRF2␤, PAC1, GLP-1, and GIP receptors and of the CRF1 or parathyroid hormone receptor N-terminal region fused with the maltose-binding protein, in complex with their respective agonist or antagonist ligands, have been determined by NMR or x-ray diffraction studies (6 -12). These structures share a common fold stabilized by three conserved disulfide bridges and by an ionic bridge involving a conserved aspartate.The scanning cysteine accessibility m...

show abstract

Section: Discussionmentioning

confidence: 99%

Mutational and Cysteine Scanning Analysis of the Glucagon Receptor N-terminal Domain

Prévost

Vertongen

Raussens

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Structural studies elucidated the folds of the CLR and RAMP1 and -2 ECDs and revealed how they interact in the absence of peptide ligands. [15][16][17] The CLR ECD fold is common to class B GPCR ECDs and consists of an N-terminal a-helix followed by a short consensus repeat fold that is composed of two b-sheets each with two b-strands. Three conserved disulfide bonds hold the secondary structure elements together.…”

mentioning

confidence: 99%

Selective CGRP and adrenomedullin peptide binding by tethered RAMP‐calcitonin receptor‐like receptor extracellular domain fusion proteins

Moad

Pioszak

2013

Protein Science

View full text Add to dashboard Cite

Calcitonin gene-related peptide (CGRP) and adrenomedullin (AM) are related peptides that are potent vasodilators. The CGRP and AM receptors are heteromeric protein complexes comprised of a shared calcitonin receptor-like receptor (CLR) subunit and a variable receptor activity modifying protein (RAMP) subunit. RAMP1 enables CGRP binding whereas RAMP2 confers AM specificity. How RAMPs determine peptide selectivity is unclear and the receptor stoichiometries are a topic of debate with evidence for 1:1, 2:2, and 2:1 CLR:RAMP stoichiometries. Here, we describe bacterial production of recombinant tethered RAMP-CLR extracellular domain (ECD) fusion proteins and biochemical characterization of their peptide binding properties. Tethering the two ECDs ensures complex stability and enforces defined stoichiometry. The RAMP1-CLR ECD fusion purified as a monomer, whereas the RAMP2-CLR ECD fusion purified as a dimer. Both proteins selectively bound their respective peptides with affinities in the low micromolar range. Truncated CGRP(27-37) and AM(37-52) fragments were identified as the minimal ECD complex binding regions. The CGRP C-terminal amide group contributed to, but was not required for, ECD binding, whereas the AM C-terminal amide group was essential for ECD binding. Alanine-scan experiments identified CGRP residues T30, V32, and F37 and AM residues P43, K46, I47, and Y52 as critical for ECD binding. Our results identify CGRP and AM determinants for receptor ECD complex binding and suggest that the CGRP receptor functions as a 1:1 heterodimer. In contrast, the AM receptor may function as a 2:2 dimer of heterodimers, although our results cannot rule out 2:1 or 1:1 stoichiometries.

show abstract

“…Smallmolecule antagonists discussed below act by blocking the binding to the peptide-binding cleft [61]. RAMP1 is involved in processing and presenting the CGRP receptor to the cell surface and helps determine the relative affinity and sensitivity of the receptor [62,63]. Zhang et al [64] showed that RAMP1 is functionally rate limiting for CGRP receptor activity in the trigeminal ganglion, suggesting that elevated RAMP1 could sensitize migraine sufferers to CGRP actions [64].…”

Section: The Receptormentioning

confidence: 99%

Targeting CGRP: A New Era for Migraine Treatment

Goldberg

Silberstein

2015

CNS Drugs

View full text Add to dashboard Cite

Migraine is a highly prevalent headache disease that typically affects patients during their most productive years. Despite significant progress in understanding the underlying pathophysiology of this disorder, its treatment so far continues to depend on drugs that, in their majority, were not specifically designed for this purpose. The neuropeptide calcitonin gene-related peptide (CGRP) has been indicated as playing a critical role in the central and peripheral pathways leading to a migraine attack. It is not surprising that drugs designed to specifically block its action are gaining remarkable attention from researchers in the field with, at least so far, a safe risk profile. In this article, we highlight the evolution from older traditional treatments to the innovative CGRP target drugs that are revolutionizing the way to approach this debilitating neurological disease. We provide a brief introduction on pathophysiology of migraine and details on the characteristic, function, and localization of CGRP to then focus on CGRP receptor antagonists (CGRP-RAs) and CGRP monoclonal antibodies (CGRP mAbs). Key PointsThe neuropeptide calcitonin gene-related peptide (CGRP) has been indicated as playing a critical role in the central and peripheral pathways leading to a migraine attack.Targeting CGRP as a specific therapeutic tool for migraine may offer similar or even better efficacy than currently available treatments without the vasoconstrictive risk factors.Further studies are necessary to determine the exact role of CGRP receptor antagonists and CGRP monoclonal antibodies in migraine with aura, allodynia, and photophobia.

show abstract

Crystal structure of the human receptor activity‐modifying protein 1 extracellular domain

Cited by 51 publications

References 37 publications

Mutational and Cysteine Scanning Analysis of the Glucagon Receptor N-terminal Domain

Mutational and Cysteine Scanning Analysis of the Glucagon Receptor N-terminal Domain

Selective CGRP and adrenomedullin peptide binding by tethered RAMP‐calcitonin receptor‐like receptor extracellular domain fusion proteins

Targeting CGRP: A New Era for Migraine Treatment

Contact Info

Product

Resources

About