Cloning, expression, and mutational analysis of the pigeon prolactin receptor.
Cloning, expression, and mutational analysis of the pigeon prolactin receptor.
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DOI：
10.1210/endo.135.1.7516866
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发表时间：
1994-07
期刊：
影响因子：
4.8
通讯作者：
X. Chen;N. Horseman
中科院分区：
文献类型：
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作者：
X. Chen;N. Horseman
通过对鸽生殖道囊文库的筛选和逆转录-聚合酶链反应的方法，获得了一条鸽催乳素受体（PRLR）的全长互补DNA。推导的830个氨基酸的序列揭示了PRL/GH/细胞因子受体超家族的一个新成员;它包含一个单一的跨膜结构域，所有保守的半胱氨酸对和WSxWS基序在其胞外结构域，和一个保守的脯氨酸丰富的基序在其胞内结构域。鸽子PRLR的胞质结构域在长度和一般序列特征上与哺乳动物PRLR的长型相似。在所检查的任何克隆中没有短或中间形式受体的证据。与哺乳动物PRLR有一个胞外结构域不同，鸽子PRLR在其胞外结构域中有两个高度同源的单元。重复单元的氨基酸序列彼此相同64%，与哺乳动物PRLR的胞外域相同52-59%（远膜单元）和60-71%（近膜单元）。为了研究重复的细胞外单位在鸽子中的意义，构建了突变的鸽子PRLR互补DNA，其在细胞外结构域中仅包含近膜单位。野生型和突变的鸽子PRLR都以同样高的亲和力（Ka = 0.6 nM-1）与大鼠PRL（rPRL）结合。两种形式的鸽子PRLR的配体特异性也是相同的。
A full-length PRL receptor (PRLR) complementary DNA from pigeons was obtained by screening pigeon crop sac libraries and by reverse transcription coupled with polymerase chain reaction. The deduced sequence of 830 amino acids revealed a new member of the PRL/GH/cytokine receptor superfamily; it contained a single transmembrane domain, all of the conserved cysteine pairs and the WSxWS motif in its extracellular domain, and a conserved proline-rich motif in its intracellular domain. The cytoplasmic domain of the pigeon PRLR was similar to the long form of mammalian PRLRs in both length and general sequence characteristics. There was no evidence of short or intermediate form receptor in any of the clones examined. Unlike mammalian PRLRs, which have a single extracellular domain, the pigeon PRLR had two highly homologous units in its extracellular domain. Amino acid sequences of the repeated units were 64% identical to each other and were 52-59% (membrane-distal unit) and 60-71% (membrane-proximal unit) identical to the extracellular domain of the mammalian PRLRs. To study the significance of the repeated extracellular units in pigeons, a mutated pigeon PRLR complementary DNA that contained only the membrane-proximal unit in the extracellular domain was constructed. Both the wild-type and mutated pigeon PRLRs bound to rat PRL (rPRL) with equally high affinities (Ka = 0.6 nM-1). The ligand specificities of both forms of the pigeon PRLR were also identical.