Identification of N-Terminal Receptor Activity-Modifying Protein Residues Important for Calcitonin Gene-Related Peptide, Adrenomedullin, and Amylin Receptor Function

Qi, Tao; Christopoulos, George; Bailey, Richard J.; Christopoulos, Arthur; Sexton, Patrick M.; Hay, Debbie L.

doi:10.1124/mol.108.047142

Cited by 66 publications

(66 citation statements)

References 29 publications

(59 reference statements)

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“…570 chimeras and site-directed mutagenesis have clearly identified that the N terminus of RAMP is important for peptide interactions (Udawela et al, 2006a,b;Qi et al, 2008). A specific RAMP1 residue (tryptophan 84) may directly contribute to amylin and CGRP interactions with the AMY 1 receptor (Gingell et al, 2010;Booe et al, 2015).…”

Section: Receptormentioning

confidence: 99%

Amylin: Pharmacology, Physiology, and Clinical Potential

Hay¹,

Chen²,

Lutz³

et al. 2015

Pharmacol Rev

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279

295

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Section: Receptormentioning

confidence: 99%

Amylin: Pharmacology, Physiology, and Clinical Potential

Hay¹,

Chen²,

Lutz³

et al. 2015

Pharmacol Rev

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279

295

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“…Both receptors also recognize CGRP with lower affinity, with the AM 2 receptor having a higher affinity compared with the AM 1 receptor, depending on the species Hay et al, 2003). It has been reported that AM2 was a nonselective agonist at the three CL/RAMP complexes (Roh et al, 2004), but recent evidence suggests that AM2 can show higher affinity for CL/RAMP3 compared with CL/RAMP1 (Qi et al, 2008). The mechanism of AM binding to its receptors is still poorly defined but is most likely to involve AM binding in a pocket formed from the extracellular N termini of CL and RAMP2 or RAMP3 (for review, see Hay et al, 2006).…”

mentioning

confidence: 99%

“…If this residue is substituted with tryptophan (found in the equivalent position of RAMP1; Trp74), AM potency and affinity are decreased and the pharmacology looks more CGRP receptor-like in profile. If the opposite substitution is made, replacing Trp74 with Glu (W74E), there is a corresponding enhancement of AM affinity, and the pharmacology looks more AM 2 receptor-like (Qi et al, 2008). CGRP potency is unaffected by these substitutions, strongly suggesting that this residue is specifically involved in AM interactions.…”

mentioning

confidence: 99%

“…The mechanism of AM binding to its receptors is still poorly defined but is most likely to involve AM binding in a pocket formed from the extracellular N termini of CL and RAMP2 or RAMP3 (for review, see Hay et al, 2006). Accumulating data suggest that glutamic acid at position 74 (Glu74) in RAMP3 may be crucial for determining high-affinity AM binding at the AM 2 receptor (Qi et al, 2008). If this residue is substituted with tryptophan (found in the equivalent position of RAMP1; Trp74), AM potency and affinity are decreased and the pharmacology looks more CGRP receptor-like in profile.…”

mentioning

confidence: 99%

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Novel Peptide Antagonists of Adrenomedullin and Calcitonin Gene-Related Peptide Receptors: Identification, Pharmacological Characterization, and Interactions with Position 74 in Receptor Activity-Modifying Protein 1/3

Robinson

Aitken²,

Bailey³

et al. 2009

J Pharmacol Exp Ther

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Human adrenomedullin (AM) is a 52-amino acid peptide belonging to the calcitonin peptide family, which also includes calcitonin gene-related peptide (CGRP) and AM2. The two AM receptors, AM 1 and AM 2 , are calcitonin receptor-like receptor (CL)/receptor activity-modifying protein (RAMP) (RAMP2 and RAMP3, respectively) heterodimers. CGRP receptors comprise CL/RAMP1. The only human AM receptor antagonist (AM ) is a truncated form of AM; it has low affinity and is only weakly selective for AM 1 over AM 2 receptors. To develop novel AM receptor antagonists, we explored the importance of different regions of AM in interactions with AM 1 , AM 2 , and CGRP receptors. AM 22-52 was the framework for generating further AM fragments (AM 26 -52 and AM 30 -52 ), novel AM/␣CGRP chimeras (C1-C5 and C9), and AM/AM 2 chimeras (C6 -C8). cAMP assays were used to screen the antagonists at all receptors to determine their affinity and selectivity. Circular dichroism spectroscopy was used to investigate the secondary structures of AM and its related peptides. The data indicate that the structures of AM, AM2, and ␣CGRP differ from one another. Our chimeric approach enabled the identification of two nonselective highaffinity antagonists of AM 1 , AM 2 , and CGRP receptors (C2 and C6), one high-affinity antagonist of AM 2 receptors (C7), and a weak antagonist selective for the CGRP receptor (C5). By use of receptor mutagenesis, we also determined that the C-terminal nine amino acids of AM seem to be responsible for its interaction with Glu74 of RAMP3. We provide new information on the structure-activity relationship of AM, ␣CGRP, and AM2 and how AM interacts with CGRP and AM 2 receptors.Human adrenomedullin (AM) is a 52-amino acid peptide belonging to the calcitonin family of peptides, which also includes calcitonin gene-related peptide (CGRP) and AM2 (also known as intermedin). The calcitonin peptide family shows weak homology at the level of the primary sequence, but there are stronger relationships at the secondary structure level. Each member of the family has an N-terminal ring structure and an amidated carboxyl terminus. Both of these structures are critical for receptor binding and subsequent signaling (Eguchi et al., 1994;Conner et al., 2002). Truncation of the N-terminal ring structure produces antagonist peptides in all family members. The ring structure of AM is created by the formation of a disulfide bond between two cysteine residues at positions 16 and 21 of the peptide (Eguchi et al., 1994). Truncation of AM yields the AM antagonist AM . Interfering with the C-terminal amidated tyrosine residue of AM results in a peptide with reduced receptor affinity (Eguchi et al., 1994). AM 13-52 and AM 15-52 are both agonists with similar affinity to full-length AM, however, little is known structurally about AM.The combination of the calcitonin receptor-like receptor (CL) with a receptor activity-modifying protein (RAMP) constitutes receptors that bind AM . CL is unusual for a G protein-coupled receptor in that it cannot f...

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“…The RAMP1/CLR combination forms the CGRP receptor, which is potently activated by CGRP; AM can also signal via this receptor, binding with around 10-fold lower affinity than CGRP , Bailey & Hay 2006. AM2 may have similar pharmacology to AM but this is poorly explored (Roh et al 2004, Takei et al 2004, Qi et al 2008. These receptors predominantly signal via activation of adenylate cyclase and increase intracellular cAMP.…”

Section: Am and Cgrp Receptorsmentioning

confidence: 99%

Adrenomedullin and calcitonin gene-related peptide receptors in endocrine-related cancers: opportunities and challenges

Hay

Walker²,

Poyner³

2010

Endocrine Related Cancer

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Adrenomedullin (AM), adrenomedullin 2 (AM2/intermedin) and calcitonin gene-related peptide (CGRP) are members of the calcitonin family of peptides. They can act as growth or survival factors for a number of tumours, including those that are endocrine-related. One mechanism through which this occurs is stimulating angiogenesis and lymphangiogenesis. AM is expressed by numerous tumour types and for some cancers, plasma AM levels can be correlated with the severity of the disease. In cancer models, lowering AM content or blocking AM receptors can reduce tumour mass. AM receptors are complexes formed between a seven transmembrane protein, calcitonin receptor-like receptor and one of the two accessory proteins, receptor activity-modifying proteins (RAMPs) 2 or 3 to give the AM 1 and AM 2 receptors respectively. AM also has affinity at the CGRP receptor, which uses RAMP1. Unfortunately, due to a lack of selective pharmacological tools or antibodies to distinguish AM and CGRP receptors, the precise receptors and signal transduction pathways used by the peptides are often uncertain. Two other membrane proteins, RDC1 and L1/G10D (the 'ADMR'), are not currently considered to be genuine CGRP or AM receptors. In order to properly evaluate whether AM or CGRP receptor inhibition has a role in cancer therapy, it is important to identify which receptors mediate the effects of these peptides. To effectively distinguish AM 1 and AM 2 receptors, selective receptor antagonists need to be developed. The development of specific CGRP receptor antagonists suggests that this is now feasible.

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Identification of N-Terminal Receptor Activity-Modifying Protein Residues Important for Calcitonin Gene-Related Peptide, Adrenomedullin, and Amylin Receptor Function

Cited by 66 publications

References 29 publications

Amylin: Pharmacology, Physiology, and Clinical Potential

Amylin: Pharmacology, Physiology, and Clinical Potential

Novel Peptide Antagonists of Adrenomedullin and Calcitonin Gene-Related Peptide Receptors: Identification, Pharmacological Characterization, and Interactions with Position 74 in Receptor Activity-Modifying Protein 1/3

Adrenomedullin and calcitonin gene-related peptide receptors in endocrine-related cancers: opportunities and challenges

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