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
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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COS Cells / metabolism
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Calcitonin Gene-Related Peptide / pharmacology
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Cells, Cultured / metabolism
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Chlorocebus aethiops
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Cloning, Molecular
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Cyclic AMP / metabolism
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Cyclic AMP-Dependent Protein Kinases / physiology
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DNA, Complementary / genetics
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HeLa Cells / drug effects
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HeLa Cells / metabolism
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Humans
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Intracellular Signaling Peptides and Proteins
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MAP Kinase Signaling System / drug effects
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Macromolecular Substances
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Membrane Proteins / genetics*
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Mice / genetics*
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Mice, Inbred C57BL
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Molecular Sequence Data
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Organ Specificity
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RNA, Messenger / biosynthesis
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RNA, Messenger / genetics
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Receptor Activity-Modifying Protein 1
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Receptor Activity-Modifying Proteins
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Receptors, Calcitonin Gene-Related Peptide / chemistry
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Receptors, Calcitonin Gene-Related Peptide / drug effects
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Receptors, Calcitonin Gene-Related Peptide / genetics*
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Recombinant Fusion Proteins / metabolism
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Second Messenger Systems / drug effects
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Sequence Alignment
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Sequence Homology, Amino Acid
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Species Specificity
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T-Lymphocytes / metabolism
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Transfection
Substances
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DNA, Complementary
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Intracellular Signaling Peptides and Proteins
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Macromolecular Substances
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Membrane Proteins
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RAMP1 protein, human
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RNA, Messenger
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Ramp1 protein, mouse
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Receptor Activity-Modifying Protein 1
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Receptor Activity-Modifying Proteins
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Receptors, Calcitonin Gene-Related Peptide
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Recombinant Fusion Proteins
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinases
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Calcitonin Gene-Related Peptide
Associated data
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GENBANK/AA015595
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GENBANK/AA019046