Molecular cloning and characterization of mouse calcitonin gene-related peptide receptor

Neuropeptides. 2002 Feb;36(1):22-33. doi: 10.1054/npep.2002.0871.

Abstract

The calcitonin gene-related peptide (CGRP) plays important roles as a neurotransmitter/neuromodulator in the central nervous system, and as a potent vasodilator when secreted from peripheral, perivascular nerves through its specific receptors. In this study, we cloned mouse cDNA counterparts of the human CGRP receptor composed of calcitonin receptor-like receptor (CRLR) and receptor activity modifying protein 1 (RAMP1) and examined the signal transduction mechanism through the CGRP receptor. Mouse CRLR (mCRLR) is a 462-amino acid G protein-coupled heptahelical receptor, and mouse RAMP1 (mRAMP1) is a 148-amino acid single membrane-spanning protein with a short cytoplasmic portion. Specific binding of (125)I-CGRP was detected only when both mCRLR and mRAMP1 cDNAs were cotransfected to COS-7 cells, and the Kd value of the receptor was 2.2 x 10(-10) M. CGRP induced a marked elevation of the intracellular cAMP levels in COS-7 cells cotransfected with mCRLR and mRAMP1. CGRP signaling through the mCRLR/mRAMP1 receptor complex was found to increase the promoter activities of cyclic AMP responsive element and serum responsive element in the co-transfected HeLa cells. These results indicate that mCRLR and mRAMP1 constitute a functional mouse CGRP receptor for the transduction of CGRP signaling by PKA and extracellular signal-regulated kinase signal transduction pathways.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • COS Cells / metabolism
  • Calcitonin Gene-Related Peptide / pharmacology
  • Cells, Cultured / metabolism
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / physiology
  • DNA, Complementary / genetics
  • HeLa Cells / drug effects
  • HeLa Cells / metabolism
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • MAP Kinase Signaling System / drug effects
  • Macromolecular Substances
  • Membrane Proteins / genetics*
  • Mice / genetics*
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Organ Specificity
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Receptor Activity-Modifying Protein 1
  • Receptor Activity-Modifying Proteins
  • Receptors, Calcitonin Gene-Related Peptide / chemistry
  • Receptors, Calcitonin Gene-Related Peptide / drug effects
  • Receptors, Calcitonin Gene-Related Peptide / genetics*
  • Recombinant Fusion Proteins / metabolism
  • Second Messenger Systems / drug effects
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Species Specificity
  • T-Lymphocytes / metabolism
  • Transfection

Substances

  • DNA, Complementary
  • Intracellular Signaling Peptides and Proteins
  • Macromolecular Substances
  • Membrane Proteins
  • RAMP1 protein, human
  • RNA, Messenger
  • Ramp1 protein, mouse
  • Receptor Activity-Modifying Protein 1
  • Receptor Activity-Modifying Proteins
  • Receptors, Calcitonin Gene-Related Peptide
  • Recombinant Fusion Proteins
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • Calcitonin Gene-Related Peptide

Associated data

  • GENBANK/AA015595
  • GENBANK/AA019046