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J Biol Chem, Vol. 274, Issue 40, 28185-28190, October 1, 1999
Zebrafish Express the Common Parathyroid Hormone/Parathyroid
Hormone-related Peptide Receptor (PTH1R) and a Novel Receptor (PTH3R)
That Is Preferentially Activated by Mammalian and Fugufish Parathyroid
Hormone-related Peptide
David A.
Rubin and
Harald
Jüppner
From the Endocrine Unit and Pediatric Services, Massachusetts
General Hospital and Harvard Medical School,
Boston, Massachusetts 02114
To further explore the evolution of receptors for
parathyroid hormone (PTH) and PTH-related peptide (PTHrP), we
searched for zebrafish (z) homologs of the PTH/PTHrP receptor (PTH1R).
In mammalian genes encoding this receptor, exons M6/7 and M7 are highly
conserved and separated by 81-84 intronic nucleotides. Genomic
polymerase chain reaction using degenerate primers based on these exons
led to two distinct DNA fragments comprising portions of genes encoding the zPTH1R and the novel zPTH3R. Sequence comparison of both
full-length teleost receptors revealed 69% similarity (61% identity),
but less homology with zPTH2R. When compared with hPTH1R, zPTH1R showed 76% and zPTH3R 67% amino acid sequence similarity; similarity with
hPTH2R was only 59% for both teleost receptors. When expressed in
COS-7 cells, zPTH1R bound
[Tyr34]hPTH-(1-34)-amide (hPTH),
[Tyr36]hPTHrP-(1-36)-amide (hPTHrP), and
[Ala29,Glu30,Ala34,Glu35, Tyr36]fugufish
PTHrP-(1-36)-amide (fuguPTHrP) with a high apparent affinity
(IC50: 1.2-3.5 nM), and was efficiently
activated by all three peptides (EC50: 1.1-1.7
nM). In contrast, zPTH3R showed higher affinity for
fuguPTHrP and hPTHrP (IC50: 2.1-11.1 nM) than for hPTH (IC50: 118.2-127.0 nM); cAMP
accumulation was more efficiently stimulated by fugufish and human
PTHrP (EC50: 0.47 ± 0.27 and 0.45 ± 0.16, respectively) than by hPTH (EC50: 9.95 ± 1.5 nM). Agonist-stimulated total inositol phosphate
accumulation was observed with zPTH1R, but not zPTH3R.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 1999 by the American Society for Biochemistry and Molecular Biology.
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