Targeting expression of the catalytic domain of the kinase insert domain receptor (KDR) in the peroxisomes of Pichia pastoris
文献类型：期刊论文
| 作者 | Wang, Ya2; Xuan, Yaoji2; Zhang, Ping2; Jiang, Xi2; Ni, Zhenhua2; Tong, Linjiang1; Zhou, Xiangshan2; Lin, Liping1; Ding, Jian1 ; Zhang, Yuanxing2
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| 刊名 | FEMS YEAST RESEARCH
 |
| 出版日期 | 2009-08 |
| 卷号 | 9期号:5页码:732-741 |
| 关键词 | KDR kinase insert domain receptor peroxisome phosphorylation Pichia pastoris vascular endothelial growth factor receptor 2 |
| ISSN号 | 1567-1356 |
| DOI | 10.1111/j.1567-1364.2009.00521.x |
| 文献子类 | Article |
| 英文摘要 | The kinase insert domain receptor (KDR), also known as vascular endothelial growth factor receptor-2 (VEGFR2), is an important therapeutic target for the treatment of cancer because of its crucial role in angiogenesis, which is fundamental to the malignancy of tumors. Here, we expressed the catalytic domain of KDR in Pichia pastoris under the control of the AOX1 promoter. In order to facilitate its purification and detection, His-tag and green fluorescent protein (GFP) were fused to the N-terminus of KDR. At the same time, a peroxisomal targeting signal 1 (SKL) was fused to the C-terminus to avoid the potential negative effect on the host cell. The highly expressing clone K1 was selected by GFP fluorescence intensity analysis using flow cytometry (FCM). Furthermore, the GFP-KDR-SKL fusion protein was proved to be correctly targeted to the peroxisomes of P. pastoris by colocation with blue fluorescent protein-SKL. The expression of GFP-KDR-SKL led to extensive phosphorylation of endogenous proteins and significantly inhibited cell growth. However, the expression was not lethal to the cells. Both in vitro biological activity assay and inhibition rate assay demonstrated that the purified GFP-KDR-SKL fusion protein exhibited high kinase catalytic activity and could be used as a target for anticancer drug screening. |
| WOS关键词 | ENDOTHELIAL GROWTH-FACTOR ; YEAST HANSENULA-POLYMORPHA ; GREEN FLUORESCENT PROTEIN ; ANTI-ANGIOGENIC AGENTS ; TYROSINE KINASE ; METHYLOTROPHIC YEAST ; CANCER-THERAPY ; RECOMBINANT PROTEIN ; GENE-EXPRESSION ; INHIBITOR |
| 资助项目 | National Basic Research (973) Program of China[2004CB518903] ; Shanghai Leading Academic Discipline Project[B505] |
| WOS研究方向 | Biotechnology & Applied Microbiology ; Microbiology ; Mycology |
| 语种 | 英语 |
| WOS记录号 | WOS:000267751800006 |
| 出版者 | OXFORD UNIV PRESS |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/279175]  |
| 专题 | 药理学第一研究室 中科院受体结构与功能重点实验室 新药研究国家重点实验室 科研与新药推进处 |
| 通讯作者 | Zhou, Xiangshan |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Mat Med, Div Antitumor Pharmacol, State Key Lab Drug Res, Shanghai 200031, Peoples R China 2.E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China; |
| 推荐引用方式 GB/T 7714 | Wang, Ya,Xuan, Yaoji,Zhang, Ping,et al. Targeting expression of the catalytic domain of the kinase insert domain receptor (KDR) in the peroxisomes of Pichia pastoris[J]. FEMS YEAST RESEARCH,2009,9(5):732-741. |
| APA | Wang, Ya.,Xuan, Yaoji.,Zhang, Ping.,Jiang, Xi.,Ni, Zhenhua.,...&Zhang, Yuanxing.(2009).Targeting expression of the catalytic domain of the kinase insert domain receptor (KDR) in the peroxisomes of Pichia pastoris.FEMS YEAST RESEARCH,9(5),732-741. |
| MLA | Wang, Ya,et al."Targeting expression of the catalytic domain of the kinase insert domain receptor (KDR) in the peroxisomes of Pichia pastoris".FEMS YEAST RESEARCH 9.5(2009):732-741. |
入库方式： OAI收割
来源：上海药物研究所
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