論文 - 詳細
| RRC ID | 38221 |
|---|---|
| 著者 | Kimura Y, Nagata K, Suzuki N, Yokoyama R, Yamanaka Y, Kitamura H, Hirano H, Ohara O. |
| タイトル | Characterization of multiple alternative forms of heterogeneous nuclear ribonucleoprotein K by phosphate-affinity electrophoresis. |
| ジャーナル | Proteomics |
| Abstract |
The phosphorylation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) is thought to play an important role in cell regulation and signal transduction. However, the relationship between hnRNP K phosphorylation and cellular events has only been indirectly examined, and the phosphorylated forms of endogenous hnRNP K have not been biochemically characterized in detail. In this study, we extensively examined the phosphorylated forms of endogenous hnRNP K by direct protein-chemical characterization using phosphate-affinity electrophoresis followed by immunoblotting and MS. Phosphate-affinity electrophoresis enabled us to sensitively detect and separate the phosphorylated forms of hnRNP K. When we used 2-DE with phosphate-affinity SDS-PAGE in the second dimension, the nuclear fraction contained more than 20 spots of endogenous hnRNP K on the 2-D map. We determined that the multiple forms of hnRNP K were produced mainly by alternative splicing of the single hnRNP K gene and phosphorylation of Ser116 and/or Ser284. Furthermore, the subcellular localization of these proteins revealed by the 2-D gel correlated with their phosphorylation states and alternative splicing patterns. The results also indicated that the multiple forms of hnRNP K were differentially modulated in response to external stimulation with bacterial lipopolysaccharide or serum. |
| 巻・号 | 10(21) |
| ページ | 3884-95 |
| 公開日 | 2010-11-1 |
| DOI | 10.1002/pmic.201000349 |
| PMID | 20960454 |
| MeSH | Alternative Splicing Animals Cell Fractionation Cell Line Cell Nucleus / chemistry Cell Nucleus / metabolism Cell-Free System Cytoplasm / chemistry Cytoplasm / metabolism Electrophoresis, Gel, Two-Dimensional / methods* Electrophoresis, Polyacrylamide Gel HeLa Cells Heterogeneous-Nuclear Ribonucleoprotein K / chemistry* Heterogeneous-Nuclear Ribonucleoprotein K / metabolism Humans Immunoblotting Lipopolysaccharides / pharmacology Mass Spectrometry Mice Phosphorylation Protein Biosynthesis / drug effects Protein Isoforms Proteomics / methods* |
| IF | 3.254 |
| 引用数 | 20 |
| WOS 分野 | BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | J774.1 |