Expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 for inhibitor screening
文献类型：期刊论文
| 作者 | Zhong, L; Guo, XN; Zhang, XH; Wu, ZX; Luo, XM ; Jiang, HL; Lin, LP; Zhang, XW; Ding, J
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| 刊名 | BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
 |
| 出版日期 | 2005-04-15 |
| 卷号 | 1722期号:3页码:254-261 |
| 关键词 | KDR bac-to-bac angiogenesis inhibitor TKI-31 |
| ISSN号 | 0304-4165 |
| DOI | 10.1016/j.bbagen.2004.12.015 |
| 文献子类 | Article |
| 英文摘要 | Vascular endothelial growth factor (VEGF), an endothelial cell-specific mitogen, can act in tumor-induced angiogenesis by binding to specific receptors on the surface of endothelial cells. One such receptor, VEGFR-2/KDR, plays a key role in VEGF-induced angiogenesis. Here, we expressed the catalytic domain of VEGFR-2 as a soluble active kinase using Bac-to-Bac expression system, and investigated correlations between VEGFR-2 activity and enzyme concentration, ATP concentration, substrate concentration and divalent cation type. We used these data to establish a convenient, effective and non-radioactive ELISA screening technique for the identification and evaluation of potential inhibitors for VEGFR-2 kinase. We screened 200 RTK target-based compounds and identified one (TKI-31) that potently inhibited VEGFR-2 kinase activity (IC50 = 0.596 mu M). Treatment of NIH3T3/KDR cells with TKI-31 blocked VEGF-induced phosphorylation of KDR in a dose-dependent manner. Moreover, TKI-31 dose-dependently suppressed HUVEC tube formation. Thus, we herein report a novel, efficient method for identifying VEGFR-2 kinase inhibitors and introduce one, TKI-31, that may prove to be a useful new angiogenesis inhibitor. (c) 2004 Elsevier B.V. All rights reserved. |
| WOS关键词 | TYROSINE KINASE DOMAIN ; INSULIN-RECEPTOR ; IN-VIVO ; ANGIOGENESIS ; IDENTIFICATION ; MECHANISM ; FLK-1 ; FLT-1 ; MICE |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000227910800003 |
| 出版者 | ELSEVIER SCIENCE BV |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/273884]  |
| 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 |
| 通讯作者 | Ding, J |
| 作者单位 | 1.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, State Key Lab Drug Res,Div Anti Tumor Pharmacol, Shanghai 201203, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, Drug Discovery & Design Ctr, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhong, L,Guo, XN,Zhang, XH,et al. Expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 for inhibitor screening[J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS,2005,1722(3):254-261. |
| APA | Zhong, L.,Guo, XN.,Zhang, XH.,Wu, ZX.,Luo, XM.,...&Ding, J.(2005).Expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 for inhibitor screening.BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS,1722(3),254-261. |
| MLA | Zhong, L,et al."Expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 for inhibitor screening".BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS 1722.3(2005):254-261. |
入库方式： OAI收割
来源：上海药物研究所
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