|
|||||
|
|
| A53Tα—synuclein与Ⅱ型单胺囊泡转移体相互作用的鉴定 | |
| 引用本文： | 陈安琪,王建英,牟青杰,刘江月,崔晓栋,张代娟,郭军堂. A53Tα—synuclein与Ⅱ型单胺囊泡转移体相互作用的鉴定[J]. 中国临床实用医学, 2008, 2(7): 43-45 |
| 作者姓名： | 陈安琪 王建英 牟青杰 刘江月 崔晓栋 张代娟 郭军堂 |
| 作者单位： | 山东省潍坊医学院,261042 |
| 摘 要： | 目的鉴定A53Tα—synuclein（A53T—SYN）与Ⅱ型囊泡单胺转移体（VMAT2）在体外分子间的相互作用，为研究α-synuelein在帕金森病（PD）发病机制中的作用提供依据。方法大肠杆菌B121中表达带GST标签的A53T-SYN蛋白，考马斯亮蓝染色和Western印迹检测蛋白质的表达，以该蛋白为诱饵，通过GST-Pull Down鉴定A53T-SYN与人脑提取物中的VMAT2在体外分子间的作用。结果考马斯亮蓝染色和Western印迹检测到GST-A53T-SYN在大肠杆菌中的高效表达，GST-Pull Down实验后的Western印迹检测发现与A53T—SYN作用的VMAT2蛋白。结论A53T-SYN与VMAT2在体外存在着分子间的相互作用。 |
| 关 键 词： | α—synuclein 帕金森氏病 Ⅱ型单胺囊泡转移体 |
Interaction between A53T mutant α-synuclein and vesicular monoamine transporter-2 |
|
| Affiliation: | CHEN An-qi, WANG Jian-ying, MU Qin-jie, et al.( Weifang Medical College, Weifang 261042, China) |
| Abstract: | Objective To identify the potential interaction between A53T mutant α-synuelein(A53TSYN)and vesicular monoamine transporter-2(VMAT2)in vitro to help studying the role of α-synuclein in Parkinson's disease.Methods CST fused A53T-SYN Were expressed in E.coli BL21 and identified by Coomassie brilliant blue R250 staining and western-blotting.GST-Pull Down experiment was used to identify the potential association between A53T-SYN and VMAT2 by incubating GST fused A53T-SYN with the extracts from norrnal human brain.Results A 40kD band of GST fused A53T-SYN was found highly expressed in E.coil BL21 by Coomassie brilliant blue B250 staining and Western-blot using an α-synuclein monoclonal antibody.A specific 62kDa band of VMAT2 was observed in the elution from the GST fused AJ3T-SYN fraction by GST-Pull Down experiment.Conclusion AS3T-SYN could interact with VMAT2 in vitro. |
| Keywords: | α-synuclein Parkinson's disease vesicular monoamine transporter-2 |
| 本文献已被 维普 万方数据 等数据库收录！ | |