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
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
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DOI：
10.1074/jbc.274.40.28185
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发表时间：
1999-10-01
影响因子：
4.8
通讯作者：
Jüppner, H
中科院分区：
文献类型：
--
作者：
Rubin, DA;Jüppner, H
为了进一步探索甲状旁腺激素（PTH）和PTH相关肽（PTHrP）受体的进化，我们寻找了PTH/PTHrP受体（PTH 1 R）的斑马鱼（z）同源物。在编码该受体的哺乳动物基因中，外显子M6/7和M7是高度保守的，并由81-84个内含子核苷酸分开。使用基于这些外显子的简并引物的基因组聚合酶链反应导致两个不同的DNA片段，其包含编码zPTH 1 R和新型zPTH 3R的基因的部分。两种受体的氨基酸序列比较显示，同源性为69%（61%），但与zPTH 2 R的同源性较低。与hPTH 1 R相比，zPTH 1 R和zPTH 3R的氨基酸序列相似性分别为76%和67%;与hPTH 2 R的氨基酸序列相似性仅为59%。当在COS-7细胞中表达时，zPTH 1 R结合[TyrS 4]hPTH-（1-34）-酰胺（hPTH）、[Tyr（36）]hPTHrP（1-36）-酰胺（hPTHrP）和[Ala（29），Glu（30），Ala（34），Glu（35），Tyr（36）]河豚PTHrP-（1-36）-酰胺（fuguPTHrP）具有高表观亲和力（IC 50：1.2-3.5 nM），并且被所有三种肽有效激活（EC 50：1.1-1.7 nM）。相比之下，zPTH 3R对fuguPTHrP和hPTHrP表现出更高的亲和力（IC50：2.1-11.1 nM）（IC 50：118.2-127.0 nM）;河豚鱼和人PTHrP更有效地刺激cAMP积累（EC 50：0.47 +/- 0.27和0.45 +/- 0.16）比hPTH（EC 50：9.95 +/- 1.5 nM）。激动剂刺激的总肌醇磷酸积累与zPTH 1 R，但不是zPTH 3R。
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 [TyrS4]hPTH-(1-34)-amide (hPTH), [Tyr(36)]hPTHrP(1-36)-amide (hPTHrP), and [Ala(29),Glu(30),Ala(34),Glu(35), Tyr(36)]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.