摘要
目的:在大肠杆菌中研究小肽促进α-突触核蛋白(α—synuclein,α—Syn)(A53T,S129A)的正常折叠。方法:利用分子生物学和生物信息学技术,以α-Syn的疏水区域为基础,设计了4种小肽(Pn:P1-P4);构建融合蛋白α-Syn—GFP,使α-Syn基因位于GFP上游,各种小肽以双顺反子方式与α-Syn—GFP共同表达;以文献报道的有效促进α-Syn正常折叠的小肽Pc(MGGAVVTGR)为对照,利用融合蛋白折叠在细菌中可视化技术研究小肽Pn影响α-Syn(A53T,S129A)的正常折叠的效果。结果:4种小肽均能更好地促进α-Syn(S129A)的正常折叠;和对照Pc相比,小肽P2(MRGGAVVTGR)使α-Syn(A53T)正常折叠提高了10%,小肽P4(MRVGGAVVR)使α-Syn(S129A)正常折叠提高了83%。结论:在大肠杆菌中,小肽可以不同程度的促进α-Syn(A53T,S129A)的正常折叠。
Objective:To study the promotion of correct folding of α-synuclein (α-Syn) ( A53T, S129A) in Escherichia coli by small peptide. Methods: The target gene was fused at upstream of GFP to give rise to α-Syn-GFP. Four small peptides ( Pn : P1 - P4) which came from the hydrophobic region of α-Syn were co-expressed with α-Syn-GFP by a hicistronic format. Using the small peptide Pc (MGGAVVTGR) as a control,the ability of the four small peptides to increase the correct folding of α-Syn (A53T, S129A) in E. coli was studied by visualization technology of fusion protein interaction in bacteria. Results: The four small peptides could increase the correct folding of α-Syn (A53T, S129A ). Small peptide P2 could increase the correct folding of α-Syn(A53T) by 10%, and P4 could increase the correct folding of α-Syn (S129A)by 83% compared with Pc. Conclusion: The design of the experiment is feasible. Small peptides can increase the correct folding of α-Syn (A53T, S129 A) in E. coll.
出处
《军事医学科学院院刊》
CSCD
北大核心
2008年第5期447-451,共5页
Bulletin of the Academy of Military Medical Sciences
基金
国家自然科学基金项目资助(30870798)
湖北工业大学校基金重大项目