Receptor activity-modifying protein-dependent impairment of calcitonin receptor splice variant Δ(1-47)hCT(a) function

T. Qi, M. Dong, H. A. Watkins, D. Wootten, Laurence J Miller, D. L. Hay

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Background and Purpose: Alternative splicing expands proteome diversity to GPCRs. Distinct receptor variants have been identified for a secretin family GPCR, the calcitonin receptor (CTR). The possible functional contributions of these receptor variants are further altered by their potential interactions with receptor activity-modifying proteins (RAMPs). One variant of the human CTR lacks the first 47 residues at its N terminus [Δ(1-47)hCT(a)]. However, very little is known about the pharmacology of this variant or its ability to interact with RAMPs to form amylin receptors. Experimental Approach: Δ(1-47)hCT(a) was characterized both with and without RAMPs in Cos7 and/or HEK293S cells. The receptor expression (ELISA assays) and function (cAMP and pERK1/2 assays) for up to six agonists and two antagonists were determined. Key Results: Despite lacking 47 residues at the N terminus, Δ(1-47)hCT(a) was still able to express at the cell surface, but displayed a generalized reduction in peptide potency. Δ(1-47)hCT (a) retained its ability to interact with RAMP1 and formed a functional amylin receptor; this also appeared to be the case with RAMP3. On the other hand, its interaction with RAMP2 and resultant amylin receptor was reduced to a greater extent. Conclusions and Implications: Δ(1-47)hCT (a) acts as a functional receptor at the cell surface. It exhibits altered receptor function, depending on whether it associates with a RAMP and which RAMP it interacts with. Therefore, the presence of this variant in tissues will potentially contribute to altered peptide binding and signalling, depending on the RAMP distribution in tissues. British Journal of Pharmacology

Original languageEnglish (US)
Pages (from-to)644-657
Number of pages14
JournalBritish Journal of Pharmacology
Volume168
Issue number3
DOIs
StatePublished - Feb 2013

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Receptor Activity-Modifying Proteins
Calcitonin Receptors
Islet Amyloid Polypeptide Receptor
Pharmacology
Peptides
Secretin
Alternative Splicing
Cell Surface Receptors
Tissue Distribution
Proteome
Enzyme-Linked Immunosorbent Assay

Keywords

  • amylin
  • calcitonin
  • G-protein coupled receptor
  • RAMP
  • receptor activity-modifying protein
  • splice variant

ASJC Scopus subject areas

  • Pharmacology

Cite this

Receptor activity-modifying protein-dependent impairment of calcitonin receptor splice variant Δ(1-47)hCT(a) function. / Qi, T.; Dong, M.; Watkins, H. A.; Wootten, D.; Miller, Laurence J; Hay, D. L.

In: British Journal of Pharmacology, Vol. 168, No. 3, 02.2013, p. 644-657.

Research output: Contribution to journalArticle

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abstract = "Background and Purpose: Alternative splicing expands proteome diversity to GPCRs. Distinct receptor variants have been identified for a secretin family GPCR, the calcitonin receptor (CTR). The possible functional contributions of these receptor variants are further altered by their potential interactions with receptor activity-modifying proteins (RAMPs). One variant of the human CTR lacks the first 47 residues at its N terminus [Δ(1-47)hCT(a)]. However, very little is known about the pharmacology of this variant or its ability to interact with RAMPs to form amylin receptors. Experimental Approach: Δ(1-47)hCT(a) was characterized both with and without RAMPs in Cos7 and/or HEK293S cells. The receptor expression (ELISA assays) and function (cAMP and pERK1/2 assays) for up to six agonists and two antagonists were determined. Key Results: Despite lacking 47 residues at the N terminus, Δ(1-47)hCT(a) was still able to express at the cell surface, but displayed a generalized reduction in peptide potency. Δ(1-47)hCT (a) retained its ability to interact with RAMP1 and formed a functional amylin receptor; this also appeared to be the case with RAMP3. On the other hand, its interaction with RAMP2 and resultant amylin receptor was reduced to a greater extent. Conclusions and Implications: Δ(1-47)hCT (a) acts as a functional receptor at the cell surface. It exhibits altered receptor function, depending on whether it associates with a RAMP and which RAMP it interacts with. Therefore, the presence of this variant in tissues will potentially contribute to altered peptide binding and signalling, depending on the RAMP distribution in tissues. British Journal of Pharmacology",
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AB - Background and Purpose: Alternative splicing expands proteome diversity to GPCRs. Distinct receptor variants have been identified for a secretin family GPCR, the calcitonin receptor (CTR). The possible functional contributions of these receptor variants are further altered by their potential interactions with receptor activity-modifying proteins (RAMPs). One variant of the human CTR lacks the first 47 residues at its N terminus [Δ(1-47)hCT(a)]. However, very little is known about the pharmacology of this variant or its ability to interact with RAMPs to form amylin receptors. Experimental Approach: Δ(1-47)hCT(a) was characterized both with and without RAMPs in Cos7 and/or HEK293S cells. The receptor expression (ELISA assays) and function (cAMP and pERK1/2 assays) for up to six agonists and two antagonists were determined. Key Results: Despite lacking 47 residues at the N terminus, Δ(1-47)hCT(a) was still able to express at the cell surface, but displayed a generalized reduction in peptide potency. Δ(1-47)hCT (a) retained its ability to interact with RAMP1 and formed a functional amylin receptor; this also appeared to be the case with RAMP3. On the other hand, its interaction with RAMP2 and resultant amylin receptor was reduced to a greater extent. Conclusions and Implications: Δ(1-47)hCT (a) acts as a functional receptor at the cell surface. It exhibits altered receptor function, depending on whether it associates with a RAMP and which RAMP it interacts with. Therefore, the presence of this variant in tissues will potentially contribute to altered peptide binding and signalling, depending on the RAMP distribution in tissues. British Journal of Pharmacology

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