論文 - 詳細
| RRC ID | 47981 |
|---|---|
| 著者 | Kinoshita E, Kinoshita-Kikuta E, Karata K, Kawano T, Nishiyama A, Yamato M, Koike T. |
| タイトル | Specific glutamic acid residues in targeted proteins induce exaggerated retardations in Phos-tag SDS-PAGE migration. |
| ジャーナル | Electrophoresis |
| Abstract |
We describe two unique proteins, Escherichia coli ClpX and human histone H2A, that show extremely retarded migrations relative to their molecular weights in Phos-tag SDS-PAGE, despite being nonphosphorylated. Although ClpX separated into multiple migration bands in Phos-tag gels, the separation was not due to phosphorylation. The N-terminal 47-61 region of ClpX was responsible for producing multiple phosphorylation-independent structural variants, even under denaturing conditions, and some of these variants were detected as highly up-shifted bands. By systematic Ala-scanning mutation analysis in the N-47-61 region, we concluded that the Glu-51 or Glu-54 residue was responsible for the appearance of exaggerated mobility-shifting bands. Histone H2A showed a much slower migration in Phos-tag gels in comparison with other major histones having similar molecular weights, and we found that the Glu-62 or Glu-65 residue caused the retarded migration. In addition, Phos-tag SDS-PAGE permitted us to detect a shift in the mobility of the phosphorylated form of histone H2A from that of the nonphosphorylated one. This is the first report showing that exaggerated retardation in the migration of a certain protein in Phos-tag SDS-PAGE is induced by interactions between the Phos-tag molecule and the carboxylate group of a specific Glu residue on the target. |
| 巻・号 | 38(8) |
| ページ | 1139-1146 |
| 公開日 | 2017-4-1 |
| DOI | 10.1002/elps.201600520 |
| PMID | 28112428 |
| MeSH | ATPases Associated with Diverse Cellular Activities Adenosine Triphosphatases Binding Sites Electrophoresis, Polyacrylamide Gel* Electrophoretic Mobility Shift Assay* Endopeptidase Clp Escherichia coli Proteins Glutamic Acid / chemistry* Histones / analysis Humans Molecular Chaperones Molecular Weight Phosphorylation Pyridines / pharmacology* |
| IF | 3.081 |
| 引用数 | 0 |
| WOS 分野 | BIOCHEMICAL RESEARCH METHODS CHEMISTRY, ANALYTICAL |
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | Genome Network Project Human cDNA W01A114G21 (HGE045765) W01A025I03 (HGE010195). |