The serine 814 of TRPC6 is phosphorylated under unstimulated conditions.
The serine 814 of TRPC6 is phosphorylated under unstimulated conditions.
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DOI：
10.1371/journal.pone.0018121
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发表时间：
2011-03-23
期刊：
影响因子：
3.7
通讯作者：
Boulay G
中科院分区：
文献类型：
--
作者：
Bousquet SM;Monet M;Boulay G
TRPC是参与钙进入的非选择性阳离子通道。它们通过磷酸化的调节已被证明可以调节它们的路径和活性。TRPC 6活性在被Fyn磷酸化后增加，并被蛋白激酶G和蛋白激酶C抑制。我们小组先前的一项研究表明，TRPC 6在人胚肾细胞系（HEK 293）中在未刺激的条件下被磷酸化。为了研究这种磷酸化的机制，我们使用了MS/MS方法结合代谢标记，并表明在位置814的丝氨酸在未刺激的细胞中被磷酸化。Ser 814突变为Ala降低了基础磷酸化，但没有改变TRPC 6活性。尽管Ser 814位于酪蛋白激酶II（CK 2）的共有位点内，但我们发现CK 2不参与TRPC 6的磷酸化，也不改变其活性。总之，我们在TRPC 6上鉴定了一个新的基础磷酸化位点（Ser 814），并表明CK 2不负责该位点的磷酸化。
TRPC are nonselective cation channels involved in calcium entry. Their regulation by phosphorylation has been shown to modulate their routing and activity. TRPC6 activity increases following phosphorylation by Fyn, and is inhibited by protein kinase G and protein kinase C. A previous study by our group showed that TRPC6 is phosphorylated under unstimulated conditions in a human embryonic kidney cells line (HEK293). To investigate the mechanism responsible for this phosphorylation, we used a MS/MS approach combined with metabolic labeling and showed that the serine at position 814 is phosphorylated in unstimulated cells. The mutation of Ser814 into Ala decreased basal phosphorylation but did not modify TRPC6 activity. Even though Ser814 is within a consensus site for casein kinase II (CK2), we showed that CK2 is not involved in the phosphorylation of TRPC6 and does not modify its activity. In summary, we identified a new basal phosphorylation site (Ser814) on TRPC6 and showed that CK2 is not responsible for the phosphorylation of this site.
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