Cloning, sequence analysis and expression of the cDNA encoding a sodium-dependent phosphate transporter from the bovine renal epithelial cell line NBL-1.
Cloning, sequence analysis and expression of the cDNA encoding a sodium-dependent phosphate transporter from the bovine renal epithelial cell line NBL-1.
复制标题
编码牛肾上皮细胞系 NBL-1 的钠依赖性磷酸转运蛋白的 cDNA 的克隆、序列分析和表达。
DOI：
10.1111/j.1432-1033.1995.0927m.x
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
J. McGivan
中科院分区：
文献类型：
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作者：
Christopher R Helps;H. Murer;J. McGivan
我们以大鼠肾脏钠依赖磷酸转运蛋白基因的一部分为探针，从ZapII NBL-1(牛肾上皮细胞)文库中克隆了一个2.2kb的全长cDNA克隆。该克隆在COS细胞系中的表达表明，它能特异编码牛肾上皮细胞的钠依赖磷酸转运蛋白。序列分析表明，该克隆有一个由693个氨基酸组成的单一开放阅读框，与大鼠和人肾脏的磷酸转运蛋白有70%的相似性[Magagnin，S.，Werner，A.，Markovich，D.，Sorrias，V.，Stange，G.，Biber，J.&Murer，H.(1993)Proc。纳特·阿卡德。SCI。美国90,5979-5983]。推导出的氨基酸序列的水化图显示了至少八个可能的跨膜区，这也与大鼠和人类转运蛋白的数据一致。该序列包含9个可能的N-糖基化位点和9个潜在的蛋白激酶C磷酸化位点。以前我们已经表明，磷向NBL-1细胞的转运动力学与负鼠肾脏细胞或大鼠肾脏的转运动力学显著不同[Help，C.&McGivan，J.(1991)EUR。J.生物化学。200、797-803]。这种差异可能与该蛋白与其他克隆的磷酸盐转运蛋白的氨基酸序列不同有关。
We have isolated a full-length cDNA clone of 2.2 kb from a lambda ZapII NBL-1 (bovine renal epithelial cell) cDNA library using a portion of the rat renal sodium-dependent phosphate transporter cDNA as a probe. Expression of this cDNA clone in the COS cell line has shown it to specifically encode a sodium-dependent phosphate transporter from bovine renal epithelial cells. Sequence analysis of the clone showed a single open reading frame of 693 amino acids which has 70% similarity to the phosphate transporter of rat and human kidney [Magagnin, S., Werner, A., Markovich, D., Sorribas, V., Stange, G., Biber, J. & Murer, H. (1993) Proc. Natl Acad. Sci. USA 90, 5979-5983]. Hydropathy plots of the derived amino acid sequence show at least eight possible transmembrane regions, again in agreement with data for the rat and human transporters. The sequence contains nine putative N-glycosylation sites and nine potential sites for protein kinase C phosphorylation. Previously we have shown that the kinetics of phosphate transport into NBL-1 cells are significantly different to those for opossum kidney cells or rat kidney [Helps, C. & McGivan, J. (1991) Eur. J. Biochem. 200, 797-803]. This difference is presumably related to differences in the amino acid sequence between this protein and other cloned phosphate transporters.