The Wilson's disease protein expressed in Sf9 cells is phosphorylated
The Wilson's disease protein expressed in Sf9 cells is phosphorylated
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DOI：
10.1042/bst0300739
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发表时间：
2002-08-01
影响因子：
3.9
通讯作者：
Lutsenko, S
中科院分区：
文献类型：
--
作者：
Vanderwerf, SM;Lutsenko, S
肝豆状核变性蛋白（WNDP）是一种铜转运蛋白，是哺乳动物细胞内铜稳态的重要介质。我们最近发现，铜浓度的变化调节WNDP的磷酸化水平。在几种哺乳动物细胞系中观察到WNDP磷酸化，表明这些细胞中存在共同的磷酸化途径。在这里，我们证明了在Sf 9昆虫细胞中表达的WNDP也被磷酸化，如通过用[P-32]P-i代谢标记这些细胞所证明的。由于杆状病毒系统允许我们产生大量的蛋白质，我们使用这种表达方法来分离WNDP并绘制WNDP磷酸化的位点。磷酸化位点的鉴定是理解WNDP磷酸化的生理作用的第一步。
The Wilson's disease protein (WNDP), a copper transporter, is a crucial mediator of copper homoeostasis in mammalian cells. We recently found that changes in copper concentration regulate the phosphorylation level of WNDP. WNDP phosphorylation was observed in several mammalian cell lines, suggesting that a common phosphorylation pathway exists in these cells. Here we demonstrate that WNDP expressed in Sf9 insect cells is also phosphorylated, as evidenced by metabolic labelling of these cells with [P-32]P-i. Because the baculovirus system allows us to generate large amounts of protein, we are using this expression method to isolate WNDP and map the sites of WNDP phosphorylation. The identification of phosphorylation sites is the first step towards understanding the physiological role of WNDP phosphorylation.