論文 - 詳細
| RRC ID | 37039 |
|---|---|
| 著者 | Nagasawa K, Meguro M, Sato K, Tanizaki Y, Nogawa-Kosaka N, Kato T. |
| タイトル | The influence of artificially introduced N-glycosylation sites on the in vitro activity of Xenopus laevis erythropoietin. |
| ジャーナル | PLoS One |
| Abstract |
Erythropoietin (EPO), the primary regulator of erythropoiesis, is a heavily glycosylated protein found in humans and several other mammals. Intriguingly, we have previously found that EPO in Xenopus laevis (xlEPO) has no N-glycosylation sites, and cross-reacts with the human EPO (huEPO) receptor despite low homology with huEPO. In this study, we introduced N-glycosylation sites into wild-type xlEPO at the positions homologous to those in huEPO, and tested whether the glycosylated mutein retained its biological activity. Seven xlEPO muteins, containing 1-3 additional N-linked carbohydrates at positions 24, 38, and/or 83, were expressed in COS-1 cells. The muteins exhibited lower secretion efficiency, higher hydrophilicity, and stronger acidic properties than the wild type. All muteins stimulated the proliferation of both cell lines, xlEPO receptor-expressing xlEPOR-FDC/P2 cells and huEPO receptor-expressing UT-7/EPO cells, in a dose-dependent manner. Thus, the muteins retained their in vitro biological activities. The maximum effect on xlEPOR-FDC/P2 proliferation was decreased by the addition of N-linked carbohydrates, but that on UT-7/EPO proliferation was not changed, indicating that the muteins act as partial agonists to the xlEPO receptor, and near-full agonists to the huEPO receptor. Hence, the EPO-EPOR binding site in X. laevis locates the distal region of artificially introduced three N-glycosylation sites, demonstrating that the vital conformation to exert biological activity is conserved between humans and X. laevis, despite the low similarity in primary structures of EPO and EPOR. |
| 巻・号 | 10(4) |
| ページ | e0124676 |
| 公開日 | 2015-1-1 |
| DOI | 10.1371/journal.pone.0124676 |
| PII | PONE-D-14-47579 |
| PMID | 25898205 |
| PMC | PMC4405594 |
| MeSH | Amino Acid Sequence Animals COS Cells Carbohydrate Conformation Cell Proliferation / drug effects Chlorocebus aethiops Dose-Response Relationship, Drug Erythropoietin / chemistry* Erythropoietin / genetics Erythropoietin / metabolism Erythropoietin / pharmacology Gene Expression Glycosylation Humans Mice Molecular Sequence Data Protein Binding Protein Engineering Receptors, Erythropoietin / genetics Receptors, Erythropoietin / metabolism* Sequence Alignment Signal Transduction Transfection Transgenes* Xenopus laevis |
| IF | 2.74 |
| 引用数 | 3 |
| WOS 分野 | 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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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 3 |
| 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 | |
| ヒト・動物細胞 | COS-1(RCB0143) |