Functional calcitonin gene-related peptide subtype 2 receptors in porcine coronary arteries are identified as calcitonin gene-related peptide subtype 1 receptors by radioligand binding and reverse transcription-polymerase chain reaction

J Pharmacol Exp Ther. 2001 Dec;299(3):1086-94.


Calcitonin gene-related peptide (CGRP) receptors are classified into CGRP subtype 1 (CGRP(1)) and CGRP subtype 2 (CGRP(2)) based on the affinity of the antagonist, human alpha (halpha)-CGRP(8-37). halpha-CGRP(8-37) antagonizes CGRP(1) receptor-mediated responses with high affinity (K(B) < 100 nM) and antagonizes CGRP(2) receptor-mediated responses with low affinity (K(B) > 1 microM). CGRP(2) receptors have been previously reported to mediate relaxation of large porcine coronary arteries because this action is antagonized with low affinity by halpha-CGRP(8-37). In the present study, we used reverse transcription-polymerase chain reaction, radioligand binding, and values from our previously reported isolated tissue experiments to compare the CGRP receptor in porcine coronary arteries with the porcine CGRP(1) receptor stably expressed in human embryonic kidney (HEK) 293 cells. We identified calcitonin receptor-like receptor and receptor activity modifying protein 1 mRNA in coronary arteries. We also found that the ligand binding characteristics of the CGRP receptor in coronary arteries and the cloned CGRP(1) receptor were highly similar. K(I) values for halpha-CGRP(8-37) were 6.6 and 5.7 nM in porcine coronary arteries and the cloned CGRP(1) receptor, respectively. The affinities (K(B)) of halpha-CGRP(8-37) and five other antagonists were 22- to 707-fold lower in functional experiments measuring relaxation of coronary arteries than in radioligand binding experiments. Despite this difference in absolute affinity values, there was a high correlation of the rank order of affinity for the antagonists determined by the two methods. Thus halpha-CGRP(8-37) antagonizes CGRP-induced relaxation of porcine coronary arteries with low affinity at the CGRP(1) receptor. Taken together, these data do not support the existence of the CGRP(2) receptor.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Calcitonin Gene-Related Peptide / metabolism
  • Calcitonin Receptor-Like Protein
  • Cells, Cultured
  • Coronary Vessels / physiology*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Iodine Radioisotopes
  • Kinetics
  • Membrane Proteins / genetics
  • Membrane Proteins / isolation & purification
  • Polymerase Chain Reaction
  • RNA, Messenger / metabolism
  • Radioligand Assay
  • Receptor Activity-Modifying Proteins
  • Receptors, Calcitonin / genetics
  • Receptors, Calcitonin / isolation & purification
  • Receptors, Calcitonin Gene-Related Peptide / physiology*
  • Swine


  • CALCRL protein, human
  • Calcitonin Receptor-Like Protein
  • Intracellular Signaling Peptides and Proteins
  • Iodine Radioisotopes
  • Membrane Proteins
  • RNA, Messenger
  • Receptor Activity-Modifying Proteins
  • Receptors, Calcitonin
  • Receptors, Calcitonin Gene-Related Peptide
  • Calcitonin Gene-Related Peptide