Protein-N-myristoylation-dependent phosphorylation of serine 13 of tyrosine kinase Lyn by casein kinase 1γ at the Golgi during intracellular protein traffic.
Protein-N-myristoylation-dependent phosphorylation of serine 13 of tyrosine kinase Lyn by casein kinase 1γ at the Golgi during intracellular protein traffic.
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DOI：
10.1038/s41598-020-73248-0
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发表时间：
2020-10-01
影响因子：
4.6
通讯作者：
Koike T
中科院分区：
文献类型：
--
作者：
Kinoshita-Kikuta E;Utsumi T;Miyazaki A;Tokumoto C;Doi K;Harada H;Kinoshita E;Koike T
Src家族激酶（SFK）的蛋白N-豆蔻酰化是将酶锚在质膜中的关键共翻译修饰。SFKs的磷酸化也是调节其酶活性的必要修饰。在这项研究中，我们使用Phos标签的SDS-PAGE研究N-豆蔻酰化依赖的SFKs及其非N-豆蔻酰化的G2 A突变体的磷酸化。林恩（Lyn-S13）的丝氨酸-13残基被证明是N-肉豆蔻酰化依赖性磷酸化。尽管已经有超过40篇关于Lyn-S13磷酸化的质谱研究报告，但负责的激酶仍然不清楚。我们成功地鉴定了酪蛋白激酶1γ（CK 1 γ）作为负责Lyn-S13磷酸化的激酶。在共表达林恩和CK 1 γ的HEK 293细胞中，Lyn-S13的磷酸化水平显著升高。CK 1 γ在CK 1家族（α、γ、δ和ε）中是独特的，携带用于膜结合的S-棕榈酰化位点。与非S-棕榈酰化CK 1 γ突变体共表达，其定位于胞质溶胶中，在Lyn-S13处的磷酸化水平没有增加。另一方面，在表达非S-棕榈酰化的Lyn-C3 A突变体的HEK 293细胞中，Lyn-C3 A突变体在Lyn-S13处被磷酸化，并且突变体保留在高尔基体处。这些结果表明，S-棕榈酰化的CK 1 γ可以在细胞内蛋白运输过程中磷酸化高尔基体上N-肉豆蔻酰化的林恩的S13。
Protein N-myristoylation of Src-family kinases (SFKs) is a critical co-translational modification to anchor the enzymes in the plasma membrane. Phosphorylation of SFKs is also an essential modification for regulating their enzymatic activities. In this study, we used Phos-tag SDS-PAGE to investigate N-myristoylation-dependent phosphorylation of SFKs and their non-N-myristoylated G2A mutants. The serine-13 residue of Lyn (Lyn-S13) was shown to be N-myristoylation-dependently phosphorylated. Although there have been more than 40 reports of mass spectrometric studies on phosphorylation at Lyn-S13, the kinase responsible remained unclear. We succeeded in identifying casein kinase 1γ (CK1γ) as the kinase responsible for phosphorylation of Lyn-S13. In HEK293 cells co-expressing Lyn and CK1γ, the phosphorylation level of Lyn-S13 increased significantly. CK1γ is unique among the CK1 family (α, γ, δ, and ε) in carrying an S-palmitoylation site for membrane binding. Co-expression with the non-S-palmitoylated CK1γ mutant, which localized in the cytosol, gave no increase in the phosphorylation level at Lyn-S13. In HEK293 cells expressing the non-S-palmitoylated Lyn-C3A mutant, on the other hand, the Lyn-C3A mutant was phosphorylated at Lyn-S13, and the mutant remained at the Golgi. These results showed that S-palmitoylated CK1γ can phosphorylate S13 of N-myristoylated Lyn at the Golgi during intracellular protein traffic.
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影响因子：
3.4
作者：
Kinoshita, Eiji;Kinoshita-Kikuta, Emiko
通讯作者：
Kinoshita-Kikuta, Emiko
影响因子：
5.8
作者：
Mitchell J;Kim SJ;Seelmann A;Veit B;Shepard B;Im E;Rhee SH
通讯作者：
Rhee SH
影响因子：
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作者：
Hu, Xiaoyi;Dang, Yongjun;Liu, Jun O.
通讯作者：
Liu, Jun O.
影响因子：
7.7
作者：
Rena, G;Bain, J;Cohen, P
通讯作者：
Cohen, P
影响因子：
7
作者：
Kinoshita, E;Kinoshita-Kikuta, E;Koike, T
通讯作者：
Koike, T