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(Received for publication, February 20, 1996) From the
Calcitonin gene-related peptide (CGRP) is a neuropeptide with
diverse biological effects including potent vasodilator activity. We
report here the cloning of a complementary DNA (cDNA) encoding a human
CGRP
Calcitonin gene-related peptide (CGRP) ( CGRP
peptides are localized predominantly in sensory afferent nerves and
central neurons(11, 12) . When released from the cell,
the peptides initiate their biological responses by binding to specific
cell surface receptors which are predominantly coupled to the
activation of adenylyl cyclase(3, 13) . CGRP receptors
have been identified and pharmacologically evaluated in several
tissues, including brain, cardiovascular, endothelial, and smooth
muscle tissues(12) . Multiple CGRP receptors have been
observed: based on pharmacological properties they are divided into at
least two subtypes and denoted as CGRP A deeper understanding of the
physiological and pathophysiological effects mediated by the CGRP
peptides has been constrained for lack of a cloned cognate receptor.
Here we describe the isolation by expressed sequence tag (EST) analysis (15, 16, 17) of a human CGRP receptor that
exhibits ligand binding and functional properties of the CGRP
We hypothesized that the CGRP and recently cloned calcitonin
(CT) receptors might display close homology since (i) like the
G-protein-coupled CT receptor(26) , CGRP signaling appears to
be mediated through the activation of adenylyl cyclase, (ii) at high
concentrations CT interacts with the CGRP binding site(27) ,
and (iii) CGRP (the Fig. 1shows the 2995-bp nucleotide sequence and deduced
protein of the cloned cDNA. Three potential in-frame ATG codons precede
the open reading frame of the protein; however, translation from the
second or third ATG codon will encode a protein with a size consistent
with that predicted for the cloned rat CT-like ``orphan''
receptor, with which it shares 91% amino acid sequence homology and
thus likely represents a receptor ortholog(18) . Also, since
the third ATG most closely approximates a Kozak consensus translation
initiation site(29) , it is probably the translation initiation
codon. Consequently, the cDNA encodes a protein of 461 amino acids,
sharing several features in common with the G-protein-linked receptors (30, 31) . Most prominent is the existence of seven
hydrophobic regions of 16 to 28 amino acids each, which are likely to
be membrane-spanning domains that form a seven-transmembrane motif
found among G-protein-coupled receptors. In addition, 52 amino acid
residues highly conserved among a recently described subfamily of
G-protein-coupled receptors including calcitonin (CT), secretin,
parathyroid, glucagon, and other
receptors(20, 26, 32, 33, 34) ,
are also present, interspersed within the sequence. Among this
subfamily, the cloned protein shares its greatest sequence identity,
55.5%, with the human CT receptor. Furthermore, within the N-terminal
domain there are several sites for post-translational modification
including three asparagine residues within consensus sites for
glycosylation and a potential cleavage site of an N-terminal
hydrophobic sequence that may be a signal peptide.
Figure 1:
Nucleotide and deduced amino acid
sequence of the human CGRP
Northern blot
analysis using the full-length cDNA as a hybridization probe revealed
mRNA species of approximately 7.5, 5.5, and 3.5 kb predominantly in the
lung and to a lesser degree in the heart (Fig. 2A). RNA
dot-blot analysis of over 20 different human tissues indicated
widespread, but generally low levels, of receptor mRNA expression (data
not shown). In situ analysis of mouse lung and heart tissues
demonstrated specific expression of the CGRP
Figure 2:
Tissue expression of CGRP
To determine the binding and functional
properties of the receptor, we used HEK 293 cells stably transfected
with the cDNA subcloned within an expression vector. Membranes prepared
from untransfected 293 cells had very little specific
Figure 3:
Pharmacological characterization of the
recombinant CGRP receptor stably expressed in 293 cells. A,
representative saturation curve for
Previous studies have shown
that CGRP mediates its responses by activation of adenylyl cyclase and
the generation of cyclic AMP(4, 5) . Treatment of
vector-transfected 293 cells with CGRP induced less than a 2-fold
increase in the accumulation of cAMP (Fig. 4A).
Addition of CGRP to 293 cells expressing the recombinant receptor
resulted in an increased accumulation of cAMP that was
concentration-dependent (Fig. 4B). The threshold,
half-maximal, and maximal concentrations of CGRP required to stimulate
cAMP accumulation in these cells were 0.1, 0.9, and 10 nM,
respectively. The maximal stimulation in response to agonist was
60-fold over basal levels of cAMP. Again, these results were comparable
to those achieved using SK-N-MC cells containing endogenous CGRP
receptors(35) . Treatment of cDNA-transfected 293 cells with
increasing concentrations of CGRP-(8-37), a selective CGRP
receptor antagonist, shifted the CGRP concentration response curve for
cAMP accumulation to the right in a parallel manner, indicating
competitive inhibition with a calculated pA
Figure 4:
Functional characterization of the
recombinant CGRP receptor. A and B,
concentration-dependent effects of CGRP (
At high concentrations, ADM evoked a specific
response (Fig. 4B), which was not surprising since this
agonist has been reported to interact weakly with the CGRP
receptor(37) . In contrast, human CT does not interact with the
recombinant receptor, since this ligand stimulated a nearly identical
activity in both CGRP receptor- and vector control-transfected cells (Fig. 4, A and B). Collectively, the
pharmacological and functional results confirm that the receptor cDNA
encodes a human CGRP Although originally
identified as an orphan rat CT-like receptor (18) and, very
recently, classified as an orphan human CT-like receptor (despite
testing for interaction with CGRP in addition to other ligands (19) ), the results presented within this report clearly
demonstrate that we have cloned a CGRP
The nucleotide sequence(s) reported in this paper has been submitted
to the GenBank(TM)/EMBL Data Bank with accession number(s)
L76380[GenBank].
Volume 271,
Number 19,
Issue of May 10, 1996 pp. 11325-11329
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES
receptor, which shares significant peptide sequence
homology with the human calcitonin receptor, a member of the
G-protein-coupled receptor superfamily. Northern blot analysis revealed
that the messenger RNA for this receptor is predominantly expressed in
the lung and heart. In situ studies showed specific
localization of the receptor mRNA to alveolar cells in the lung and to
cardiac myocytes in the heart. Stable expression of the cDNA in human
embryonic kidney 293 (HEK 293) cells produced specific, high affinity
binding sites for CGRP that displayed pharmacological and functional
properties very similar to native human CGRP receptor.
Exposure of these cells to CGRP resulted in a 60-fold increase in cAMP
production, which was inhibited in a competitive manner by the
CGRP receptor antagonist, CGRP-(8-37).
)is 37-amino
acid peptide that exists as highly homologous 
or 
isoforms
in both human and rat (1, 2) . - and -CGRP
display very similar biological activities, including peripheral and
cerebral vasodilation(3) , cardiac acceleration(4) ,
regulation of calcium metabolism(5) , reduction of intestinal
motility(6) , regulation of glucose metabolism (reduction of
insulin secretion and insulin sensitivity)(7) , diminution of
appetite (8) , and reduction of growth hormone
release(9) . The two CGRP peptides differ by 3 amino acids in
humans and 1 amino acid in rats. The amino acid sequences of CGRP
peptides are well conserved among species and can be considered as
members of a family of peptides including the related peptides amylin
(46% homology), salmon calcitonin (32% homology), and adrenomedullin
(24% homology). These peptides in general have N-terminal ring
structures of 6-7 amino acids involving a disulfide bridge and an
amidated C-terminal end(10, 11, 12) . and
CGRP
, according to the classification of Dennis et
al.(14) . CGRP-(8-37), which lacks 7 N-terminal
amino acid residues, is a selective antagonist of CGRP receptors, whereas the linear analog of CGRP, diacetoamidomethyl
cysteine CGRP (Cys(ACM2,7)CGRP), is a selective agonist of CGRP receptors(14) . receptor. Curiously, this receptor had been cloned previously as
an orphan calcitonin-like receptor receptor (rat (18) and,
recently, human(19) ), but the authors had been unable to
demonstrate an interaction with CGRP or any other ligands.
Materials
[2-[
I]iodohistidyl
]Human
(h) CGRP (specific activity 2000 Ci/mmol) was obtained from
Amersham. hCGRP, hCGRP-(8-37), salmon calcitonin (CT),
human CT, porcine vasoactive intestinal peptide, and angiotensin II
were purchased from Bachem Biochemicals. Cys(ACM2,7)CGRP was obtained
from Phoenix Pharmaceuticals (Belmont, CA). Salmon CT-(8-32) was
synthesized at SmithKline Beecham Pharmaceuticals. BCA protein assay
kit was obtained from Pierce. All other reagents were obtained from
Sigma.cDNA Cloning
Expressed sequence tag (EST) analysis (15, 16, 17) of cDNA clones derived from a
human synovial tissue cDNA library (oligo(dT)-primed and constructed in
the 
ZAPII vector (Stratagene)) identified a 800-bp clone
demonstrating significant homology to the human CT receptor. This cDNA
clone encoding an incomplete hCGRP receptor was used as a
probe to screen an oligo(dT)-primed human lung cDNA library constructed
in the ZAPII vector (Stratagene). cDNA library construction and
screening were carried out essentially as described(20) .
Several positive clones were obtained, the longest of which was
sequenced to completion by a ABI sequencer (15) (GenBank
accession number L76380).Stable Expression in HEK 293 Cells
The CGRP
receptor has three potential in-frame ATG start codons; however, the
most 3[prime-ATG codon resides in the most favorable Kozak
consensus context. For this reason, and to potentially increase protein
translation efficiency(21) , we prepared a 1.4-kilobase cDNA
fragment by PCR amplification that encompassed the entire CGRP receptor
coding region, beginning with the 3`-most ATG codon, and subcloned this
fragment into the mammalian expression vector, pCDN(22) . The
oligonucleotide primers used for PCR amplification were 5`-G GGG TAC
CCC ACC ATG GAG AAA AAG TGT ACC TCG TAT TTT CTG G-3` and 5`-CGG GAT CCC
GCA AAC AGT GAG ACA ACC ATC CTT CTA TTT TCA AT-3` (the translation
start and stop codons are underlined). Human HEK 293 cells were grown
in 100-mm culture dishes and transfected with 10 µg of the
CDN-CGRP-receptor cDNA using the LipofectAMINE transfection reagent
(Life Technologies, Inc.) according to the manufacturer's
instructions. After 2 weeks of G418 selection (0.8 mg/ml), colonies
were picked and expanded. A single cell line (among many potential
candidates) that functionally responded to CGRP treatment (by the
generation of cAMP) was chosen for further analysis. A single cell line
transfected with pCDN vector alone was selected for use as a control
for functional and binding assays.Northern Blot Analysis
Total RNA was isolated from
human tissues by TriZOL Reagent (Life Technologies, Inc.). 20 µg of
total RNA from each tissue was separated by formaldehyde, 1.0% agarose
gel electrophoresis and transferred to a nylon membrane. The
full-length human CGRP cDNA was labeled and used as probe.
Hybridization was done essentially as described(23) .In Situ Gene Amplification Analysis
Under
anesthesia, a BALB/C mouse was perfused with 4%
paraformaldehyde/phosphate-buffered saline, pH 7.5. Organs were
dissected, fixed in 10% buffered formalin at room temperature for 3
days, then embedded in paraffin. Sections of 5 µm were prepared.
Reverse transcriptase in situ gene amplification was performed
as described(24) . Two primers used in this study were chosen
from regions unique to the human CGRP and very similar to
the rat CT-like cDNA sequence (18) receptor, but not
significantly conserved in the calcitonin receptor(20) : the
upstream primer sequence was 5` GAC ATC CAG CAA GCA ACA GA 3`; the
downstream primer sequence was 5` CA ATG CCA AGC AAT GGC ACC 3`.
Digoxigenin-dUTP was applied to the reaction of gene amplification. The
products derived from gene amplification were detected by
anti-digoxigenin antibody conjugated to alkaline phosphatase (Genius3,
Boehringer Mannheim).Binding Assays
Radioligand binding assays were
performed as described(25) . In saturation binding studies,
increasing concentrations of I-CGRP (8 pM to 150
pM) were added to membranes (
25 µg) and incubated in
a total volume of 500 µl for 60 min, at 25 °C. In competition
binding studies, the membranes (30 µg of membrane proteins)
were incubated with increasing concentrations (1 pM to 1
µM) of competing ligands and 60 pMI-CGRP for 60 min at 25 °C. Nonspecific binding
was defined in the presence of 1 µM CGRP and was
usually 20% for transfected cell membranes and <10% for human
neuroblastoma SK-N-MC cell (ATCC HTB-10) membranes. No specific binding
was observed in membranes of untransfected control 293 cells.Functional Assays
Transformed or untransformed 293
cells were plated at 2.5 
10
cells/well in 6-well
plates. On day 4, the medium was aspirated and the cells were washed
with 1 ml of Dulbecco's phosphate-buffered saline containing 0.5
mM isobutylmethylxanthine for 10 min at room temperature. The
cells were treated with various concentrations (1 pM to 1
µM) of CGRP or related peptides at 37 °C for 10
min. The reaction was stopped by addition of 100 µl of 100%
ice-cold trichloroacetic acid to each well, and cAMP in each well was
measured following the radioimmunoassay protocol as described (Advance
Magnetics). Each experiment was performed in triplicate and repeated
2-3 times with different passages of cells.
, not , isopeptide) and CT share the same
mRNA transcript and are produced through alternate
splicing(1) , as are the substance P/K ligands that
cross-interact with tachykinin receptors (28) . Thus, when
expressed sequence tag (EST) analysis (15, 16, 17) of cDNA clones derived from a
human synovial tissue cDNA library identified a clone demonstrating
significant homology to the human CT receptor, this cDNA was selected
for further evaluation. Since the cDNA insert was incomplete (i.e. about 800 bp), it was used as a probe to screen a human lung cDNA
library to isolate a clone with a complete open reading frame.
receptor. Amino acids
(represented by the one-letter code) are indicated below their
respective codons and numbered 1 at the left beginning with
the ATG initiation methionine codon (M). Other numbers on the right indicate nucleotide positions starting at position 1 of
the A nucleotide of the ATG codon. Underlined amino acid
sequences, putative seven transmembrane domains; potential sites for
post-translational glycosylation are marked above the sequence by closed circle; circled amino acids are those that are
highly conserved among the secretin/CT/PTH/PTH-RP/GLP1/GHRH/GLU
G-protein-coupled receptor
subfamily(20, 26, 30, 31, 32) ; arrow, a putative secretory signal sequence cleavage site; double-underlined ATG codons, in-frame potential initiation
codons; underlined AATAAA sequence, consensus polyadenylation
signal.
receptor mRNA
in alveolar cells in the lung and cardiac myocytes in the heart (Fig. 2B).
receptor mRNA. A, Northern blot analysis of CGRP mRNA expression. RNA size markers are indicated on the left.
B, in situ gene amplification analysis of CGRP mRNA expression. The adjacent sections of mouse lung (a, b, c, d, 100) and heart (e, f, g, h, 400) were used for these
studies. a and e, use of the reverse transcriptase. b and f, omission of the reverse transcriptase. c and g, omission of amplification primers. d and h, hematoxylin and eosin staining.
I-CGRP binding (data not shown). Membranes prepared from
transfected 293 cells displayed high-affinity, low-density binding
sites for I-CGRP. The apparent dissociation constant (K
) and maximum binding (B
)
were 19 ± 3 pM and 86 ± 14 fmol/mg protein,
respectively (Fig. 3A). In competition binding studies,
the rank order potency for a series of related peptides to inhibit I-CGRP binding to these membranes was CGRP >
CGRP-(8-37) > Cys(ACM2,7)CGRP > adrenomedullin (ADM) >
salmon CT-(8-32) >>> salmon CT > human CT >
vasoactive intestinal peptide > angiotensin II (Fig. 3B). The binding affinity of I-CGRP
for the recombinant receptor as well as the pharmacological profile of
the competing ligands were very similar to that observed for endogenous
CGRP receptors present in membranes prepared from human neuroblastoma
SK-N-MC cells (Fig. 3C).
I-CGRP binding to the
membranes prepared from 293 cells expressing the recombinant CGRP
receptor. 
, total binding; 
, nonspecific binding; and
, specific binding. Inset, corresponding Scatchard plot
of the data. B, competition binding profile of CGRP (
)
and its analogs CGRP-(8-37) (), Cys(ACM2,7)CGRP (
),
ADM (
), salmon CT (), salmon CT-(8-32) (),
human CT (
), angiotensin II (
), and vasoactive intestinal
peptide (
) for specific I-CGRP binding to the
membranes. C, competition binding profile of CGRP and its
analogs for specific I-CGRP binding to SK-N-MC cell
membranes.
value of 7.57 (Fig. 4C). Except at high concentrations (1
µM), the CGRP receptor-selective agonist
Cys(ACM2,7)CGRP failed to stimulate cAMP in both receptor-transfected
cells (Fig. 4B) and SK-N-MC cells (data not shown). The
effect of Cys(ACM2,7)CGRP on the recombinant CGRP receptor in
inhibiting only I CGRP binding but not a functional cAMP
response to CGRP is consistent with the properties of the native rat
CGRP receptor present in spleen membranes(36) .
Thus, the receptor described in the present study is a CGRP receptor.
), ADM (), human
CT (), and Cys(ACM2,7)CGRP (
) on cAMP accumulation in CDN
vector-transfected 293 cells (A) and CGRP receptor-transfected
293 cells (B). C, effect of CGRP-(8-37) on
CGRP-mediated cAMP response in the absence or presence of 100 or 300
nM CGRP-(8-37). The antagonist effect was studied by
performing CGRP concentration response curves in the absence ()
and presence of 100 () and 300 () nM CGRP-(8-37).
receptor that is functionally coupled
to the activation of adenylyl cyclase. receptor. As to the
reason why previous investigators were unable to identify the reported
CT-like receptor as the CGRP receptor is only a matter of
speculation. It is possible that the recombinant CGRP receptor can be
functionally expressed only in certain cell types. In this study we
used HEK 293 cells, whereas in other reports it appears that only COS
and OK cells were used for receptor evaluation. We have recently cloned
the porcine CGRP receptor ortholog of the human receptor
and found it to exhibit essentially identical pharmacological
properties, which provides further supporting evidence that we have
identified the CGRP receptor. ()The availability
of the CGRP receptor should facilitate the study of the
physiology and pathophysiology of CGRP as a neurotransmitter,
neuromodulator, local hormone, and inflammatory mediator.
))
We thank the following for their support of the work:
William Haseltine, Craig Rosen, Michael Antonaccio, Pat Dillon,
Gou-Liang Yu, and Lily Xing at Human Genome Sciences, the sequencing
group at The Institute for Genomic Research, Martin Rosenberg, Robert
Ruffolo, Jr., Christine Debouck, Giora Feurstein, Jeff Stadel, George
Livi, Ganesh Sathe, and Subinay Ganguly at SmithKline Beecham
Pharmaceuticals.
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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 M. T. Tran, M. H. Ritchie, R. N. Lausch, and J. E. Oakes Calcitonin Gene-Related Peptide Induces IL-8 Synthesis in Human Corneal Epithelial Cells J. Immunol., April 15, 2000; 164(8): 4307 - 4312. [Abstract] [Full Text] [PDF]
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 J. P. Hinson, S. Kapas, and D. M. Smith Adrenomedullin, a Multifunctional Regulatory Peptide Endocr. Rev., April 1, 2000; 21(2): 138 - 167. [Abstract] [Full Text]
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 M. I. Rosenblatt, G. P. Dahl, and I. M. Dickerson Characterization and Localization of the Rabbit Ocular Calcitonin Gene-Related Peptide (CGRP)-Receptor Component Protein (RCP) Invest. Ophthalmol. Vis. Sci., April 1, 2000; 41(5): 1159 - 1167. [Abstract] [Full Text]
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J. A. Carucci, R. Ignatius, Y. Wei, A. M. Cypess, D. A. Schaer, M. Pope, R. M. Steinman, and S. Mojsov Calcitonin Gene-Related Peptide Decreases Expression of HLA-DR and CD86 by Human Dendritic Cells and Dampens Dendritic Cell-Driven T Cell-Proliferative Responses Via the Type I Calcitonin Gene-Related Peptide Receptor J. Immunol., April 1, 2000; 164(7): 3494 - 3499. [Abstract] [Full Text] [PDF]
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A. Martinez, S. Kapas, M.-J. Miller, Y. Ward, and F. Cuttitta Coexpression of Receptors for Adrenomedullin, Calcitonin Gene-Related Peptide, and Amylin in Pancreatic {beta}-Cells Endocrinology, January 1, 2000; 141(1): 406 - 411. [Abstract] [Full Text] [PDF]
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S. Y. K. Yousufzai, N. Ali, and A. A. Abdel-Latif Effects of Adrenomedullin on Cyclic AMP Formation and on Relaxation in Iris Sphincter Smooth Muscle Invest. Ophthalmol. Vis. Sci., December 1, 1999; 40(13): 3245 - 3253. [Abstract] [Full Text] [PDF]
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 H. Yang, K. Kawakubo, and Y. Tache Intracisternal PYY increases gastric mucosal resistance: role of cholinergic, CGRP, and NO pathways Am J Physiol Gastrointest Liver Physiol, September 1, 1999; 277(3): G555 - G562. [Abstract] [Full Text] [PDF]
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N. Bühlmann, K. Leuthäuser, R. Muff, J. A. Fischer, and W. Born A Receptor Activity Modifying Protein (RAMP)2-Dependent Adrenomedullin Receptor Is a Calcitonin Gene-Related Peptide Receptor when Coexpressed with Human RAMP1 Endocrinology, June 1, 1999; 140(6): 2883 - 2890. [Abstract] [Full Text]
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N. J. Fraser, A. Wise, J. Brown, L. M. McLatchie, M. J. Main, and S. M. Foord The Amino Terminus of Receptor Activity Modifying Proteins Is a Critical Determinant of Glycosylation State and Ligand Binding of Calcitonin Receptor-Like Receptor Mol. Pharmacol., June 1, 1999; 55(6): 1054 - 1059. [Abstract] [Full Text]
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W. Balkan, E. L. Oates, G. A. Howard, and B. A. Roos Testes Exhibit Elevated Expression of Calcitonin Gene-Related Peptide Receptor Component Protein Endocrinology, March 1, 1999; 140(3): 1459 - 1469. [Abstract] [Full Text]
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J.-C. Marie, A. Wakkach, A.-M. Coudray, E. Chastre, S. Berrih-Aknin, and C. Gespach Functional Expression of Receptors for Calcitonin Gene-Related Peptide, Calcitonin, and Vasoactive Intestinal Peptide in the Human Thymus and Thymomas from Myasthenia Gravis Patients J. Immunol., February 15, 1999; 162(4): 2103 - 2112. [Abstract] [Full Text] [PDF]
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M.-H. Huang, P. R. Knight III, and J. L. Izzo Jr. Ca2+-induced Ca2+ release involved in positive inotropic effect mediated by CGRP in ventricular myocytes Am J Physiol Regulatory Integrative Comp Physiol, January 1, 1999; 276(1): R259 - R264. [Abstract] [Full Text] [PDF]
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H. Drissi, F. Lasmoles, V. Le Mellay, P. J. Marie, and M. Lieberherr Activation of Phospholipase C-beta 1 via Galpha q/11 during Calcium Mobilization by Calcitonin Gene-related Peptide J. Biol. Chem., August 7, 1998; 273(32): 20168 - 20174. [Abstract] [Full Text] [PDF]
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H. Kaneko, T. Mitsuma, H. Nagai, S. Mori, T. Iyo, K. Kusugami, and Y. Tache Central action of adrenomedullin to prevent ethanolinduced gastric injury through vagal pathways in rats Am J Physiol Regulatory Integrative Comp Physiol, June 1, 1998; 274(6): R1783 - R1788. [Abstract] [Full Text] [PDF]
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N. A. Elshourbagy, J. E. Adamou, A. M. Swift, J. Disa, J. Mao, S. Ganguly, D. J. Bergsma, and N. Aiyar Molecular Cloning and Characterization of the Porcine Calcitonin Gene-Related Peptide Receptor Endocrinology, April 1, 1998; 139(4): 1678 - 1683. [Abstract] [Full Text] [PDF]
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Y. Isumi, H. Shoji, S. Sugo, T. Tochimoto, M. Yoshioka, K. Kangawa, H. Matsuo, and N. Minamino Regulation of Adrenomedullin Production in Rat Endothelial Cells Endocrinology, March 1, 1998; 139(3): 838 - 846. [Abstract] [Full Text] [PDF]
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 T. Tsuruda, J. Kato, K. Kitamura, K. Kuwasako, T. Imamura, Y. Koiwaya, T. Tsuji, K. Kangawa, and T. Eto Adrenomedullin: A Possible Autocrine or Paracrine Inhibitor of Hypertrophy of Cardiomyocytes Hypertension, January 1, 1998; 31(1): 505 - 510. [Abstract] [Full Text] [PDF]
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M. Naghashpour, M. I. Rosenblatt, I. M. Dickerson, and G. P. Dahl Inhibitory Effect of Calcitonin Gene-Related Peptide on Myometrial Contractility Is Diminished at Parturition Endocrinology, October 1, 1997; 138(10): 4207 - 4214. [Abstract] [Full Text] [PDF]
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V. MartInez, F. Cuttitta, and Y. Tache Central Action of Adrenomedullin to Inhibit Gastric Emptying in Rats Endocrinology, September 1, 1997; 138(9): 3749 - 3755. [Abstract] [Full Text] [PDF]
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P. D. Upton, C. Austin, G. M. Taylor, K. A. Nandha, A. J. L. Clark, M. A. Ghatei, S. R. Bloom, and D. M. Smith Expression of Adrenomedullin (ADM) and Its Binding Sites in the Rat Uterus: Increased Number of Binding Sites and ADM Messenger Ribonucleic Acid in 20-Day Pregnant Rats Compared with Nonpregnant Rats Endocrinology, June 1, 1997; 138(6): 2508 - 2514. [Abstract] [Full Text] [PDF]
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L. J. Deftos, MD Editorial: There's Something Fishy and Perhaps Even Fowl about the Mammalian Calcitonin Receptor and Its Ligand Endocrinology, February 1, 1997; 138(2): 519 - 520. [Full Text] [PDF]
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J.-F. Shyu, D. Inoue, R. Baron, and W. C. Horne The Deletion of 14Amino Acids in the Seventh Transmembrane Domain of a Naturally Occurring Calcitonin Receptor Isoform Alters Ligand Binding and Selectively Abolishes Coupling to Phospholipase C J. Biol. Chem., December 6, 1996; 271(49): 31127 - 31134. [Abstract] [Full Text] [PDF]
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K. Kuwasako, Y. Shimekake, M. Masuda, K. Nakahara, T. Yoshida, M. Kitaura, K. Kitamura, T. Eto, and T. Sakata Visualization of the Calcitonin Receptor-like Receptor and Its Receptor Activity-modifying Proteins during Internalization and Recycling J. Biol. Chem., September 15, 2000; 275(38): 29602 - 29609. [Abstract] [Full Text] [PDF]
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B. N. Evans, M. I. Rosenblatt, L. O. Mnayer, K. R. Oliver, and I. M. Dickerson CGRP-RCP, a Novel Protein Required for Signal Transduction at Calcitonin Gene-related Peptide and Adrenomedullin Receptors J. Biol. Chem., September 29, 2000; 275(40): 31438 - 31443. [Abstract] [Full Text] [PDF]
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K. Kuwasako, K. Kitamura, K. Ito, T. Uemura, Y. Yanagita, J. Kato, T. Sakata, and T. Eto The Seven Amino Acids of Human RAMP2 (86) and RAMP3 (59) Are Critical for Agonist Binding to Human Adrenomedullin Receptors J. Biol. Chem., December 21, 2001; 276(52): 49459 - 49465. [Abstract] [Full Text] [PDF]
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I. Millet, R. J. Phillips, R. S. Sherwin, S. Ghosh, R. E. Voll, R. A. Flavell, A. Vignery, and M. Rincon Inhibition of NF-kappa B Activity and Enhancement of Apoptosis by the Neuropeptide Calcitonin Gene-related Peptide J. Biol. Chem., May 12, 2000; 275(20): 15114 - 15121. [Abstract] [Full Text] [PDF]
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