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大鼠胎盘催乳素(PRL)家族新成员催乳素样蛋白D(PLP-D)的分子克隆与特性分析

Molecular cloning and characterization of a new member of the rat placental prolactin (PRL) family, PRL-like protein D (PLP-D).

作者信息

Iwatsuki K, Shinozaki M, Hattori N, Hirasawa K, Itagaki S, Shiota K, Ogawa T

机构信息

Laboratory of Cellular Biochemistry, University of Tokyo, Japan.

出版信息

Endocrinology. 1996 Sep;137(9):3849-55. doi: 10.1210/endo.137.9.8756556.

Abstract

The rat placental PRL family consists of molecules structurally similar to PRL and GH, and to date, seven members have been identified. During investigation of pregnancy stage-specific placental factors by the differential display method, we obtained a complementary DNA (cDNA) fragment (199 bp) encoding a peptide homologous to PRL-like protein (PLP)-C. By using the 3' and 5' rapid amplification of cDNA ends method, a full-length cDNA was cloned and tentatively named PLP-D. The cDNA encoded a mature protein of 240 amino acids, including a 29-amino acid signal sequence. PLP-D contains one putative N-glycosylation site and six cysteine residues that are highly conserved in the placental PRL family. Sequence comparison between PLP-D and other members of the placental PRL family showed that PLP-D is highly homologous to PLP-C (80%) and decidual PRL-related protein (73%). Northern blot analysis revealed that PLP-D messenger RNA (mRNA) first appeared at day 14 of pregnancy, and that its expression increased until term. In situ hybridization analysis indicated that PLP-D mRNA was specifically expressed in spongiotrophoblast cells and trophoblast giant cells of the placental junctional zone. Differentiated Rcho-1 cells also expressed PLP-D mRNA, whereas undifferentiated Rcho-1 cells did not.

摘要

大鼠胎盘催乳素(PRL)家族由结构上与PRL和生长激素(GH)相似的分子组成,迄今为止,已鉴定出七个成员。在通过差异显示法研究妊娠阶段特异性胎盘因子的过程中,我们获得了一个编码与催乳素样蛋白(PLP)-C同源肽的互补DNA(cDNA)片段(199 bp)。通过使用cDNA末端快速扩增3'和5'法,克隆了一个全长cDNA,并暂定名为PLP-D。该cDNA编码一个由240个氨基酸组成的成熟蛋白,包括一个29个氨基酸的信号序列。PLP-D包含一个假定的N-糖基化位点和六个半胱氨酸残基,这些残基在胎盘PRL家族中高度保守。PLP-D与胎盘PRL家族其他成员之间的序列比较表明,PLP-D与PLP-C高度同源(80%),与蜕膜PRL相关蛋白高度同源(73%)。Northern印迹分析显示,PLP-D信使核糖核酸(mRNA)在妊娠第14天首次出现,其表达在足月前增加。原位杂交分析表明,PLP-D mRNA在胎盘交界区的海绵滋养层细胞和滋养层巨细胞中特异性表达。分化的Rcho-1细胞也表达PLP-D mRNA,而未分化的Rcho-1细胞则不表达。

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