CAJ | 학술논문

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-synuclein在帕金森病(PD)发病机制中的作用提供依据.方法大肠杆菌BL21中表达带GST标签的A53T-SYN蛋白,考马斯亮蓝染色和Westem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.
목적 감정A53Tα-synuclein(A53T-SYN)여Ⅱ형낭포단알전이체(VMAT2)재체외분자간적상호작용,위연구α-synuclein재파금삼병(PD)발병궤제중적작용제공의거.방법 대장간균BL21중표체대GST표첨적A53T-SYN단백,고마사량람염색화Westem인적검측단백질적표체,이해단백위유이,통과GST-Pull Down감정A53T-SYN여인뇌제취물중적VMAT2재체외분자간적작용.결과 고마사량람염색화Western인적검측도GST-A53T-SYN재대장간균중적고효표체.GST-Pull Down실험후적Western인적검측발현여A53T-SYN작용적VMAT2단백.결론 A53T-SYN여VMAT2재체외존재착분자간적상호작용.
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.