Protein-N-myristoylation-dependent phosphorylation of serine 13 of tyrosine kinase Lyn by casein kinase 1γ at the Golgi during intracellular protein traffic
| Autor: | Haruna Harada, Tohru Koike, Aya Miyazaki, Toshihiko Utsumi, Emiko Kinoshita-Kikuta, Kyosuke Doi, Eiji Kinoshita, Chiharu Tokumoto |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell biology Science Golgi Apparatus environment and public health Biochemistry Article Mass Spectrometry Serine 03 medical and health sciences symbols.namesake 0302 clinical medicine LYN hemic and lymphatic diseases Humans Phosphorylation Multidisciplinary Cell-Free System Chemistry Kinase Casein Kinase I Biological techniques hemic and immune systems Golgi apparatus Protein-Tyrosine Kinases Cytosol Protein Transport enzymes and coenzymes (carbohydrates) 030104 developmental biology HEK293 Cells Microscopy Fluorescence 030220 oncology & carcinogenesis symbols Medicine Electrophoresis Polyacrylamide Gel lipids (amino acids peptides and proteins) Casein kinase 1 Tyrosine kinase |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-13 (2020) |
| ISSN: | 2045-2322 |
| Popis: | 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. This work was supported in part by KAKENHI Grants 18K065960 to E.K.-K., 19K071470 to E.K., and 17K08237 to T.K., and by a research grant from Chugoku Regional Innovation Research Center to E.K. |
| Databáze: | OpenAIRE |
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