[1]安 颖),吕 杰),张迎娜),等.重症肌无力患者肌细胞中LRP4胞内区与SNX17的相互作用*[J].郑州大学学报(医学版),2017,(02):129-134.[doi:10.13705/j.issn.1671-6825.2017.02.006]
 AN Ying),LYU Jie),ZHANG Yingna),et al.Expression of LRP4 intracellular region and its interaction with SNX17 in muscle cells from patients with myasthenia gravis[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2017,(02):129-134.[doi:10.13705/j.issn.1671-6825.2017.02.006]
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重症肌无力患者肌细胞中LRP4胞内区与SNX17的相互作用*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2017年02期
页码:
129-134
栏目:
论著
出版日期:
2017-03-15

文章信息/Info

Title:
Expression of LRP4 intracellular region and its interaction with SNX17 in muscle cells from patients with myasthenia gravis
作者:
安 颖1)吕 杰2)张迎娜2)高 峰2)#张 婧2)方 华2)李倩如3)杜 英3)张清勇4)王金兰1)#张运克5)
1)郑州大学第二附属医院神经内科 郑州 450014
2)郑州大学医药科学研究院神经免疫学研究室 郑州 450052
3)郑州大学基础医学院免疫学系 郑州 450001
4)郑州大学第二附属医院普胸外科 郑州 450014
5)河南中医药大学康复医学院康复办公室 郑州 450000
Author(s):
AN Ying1)LYU Jie2)ZHANG Yingna2)GAO Feng2)ZHANG Jing2)FANG Hua2)LI Qianru3)DU Ying3)ZHANG Qingyong4)WANG Jinlan1)ZHANG Yunke5)
1)Department of Neurology, the Second Affiliated Hospital, Zhengzhou University, Zhengzhou 450014
2)Department of Neuroimmunology, Institute of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052
3)Department of Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001
4)Department of Thoracic Surgery, the Second Affiliated Hospital,Zhengzhou University, Zhengzhou 450014
5)Department of Rehabilitation,Institute of Rehabilitation Medicine, Henan University of Traditional Chinese Medicine, Zhengzhou 450000
关键词:
低密度脂蛋白受体相关蛋白4 分拣微管连接蛋白17 重症肌无力 免疫共沉淀
Keywords:
low density lipoprotein receptor-related protein 4 sorting nexin 17 myasthenia gravis co-immunoprecipitation
分类号:
R746.1
DOI:
10.13705/j.issn.1671-6825.2017.02.006
摘要:
目的:探讨低密度脂蛋白受体相关蛋白4(LRP4)和分拣微管连接蛋白17(SNX17)在重症肌无力(MG)患者肌细胞中相互作用关系。方法:分别构建原核表达载体pET30a-LRP4胞内区和真核表达载体pEASY-Blunt M2-SNX17,在大肠杆菌BL21(DE3)中诱导表达相对分子质量约为17 800的His-LRP4胞内区,在HEK293T细胞中诱导表达相对分子质量约为53 000的Myc-SNX17。利用抗Myc单克隆抗体磁珠与抗His单克隆抗体磁珠进行双向免疫共沉淀(Co-IP),以验证LRP4、SNX17两种蛋白分子是否存在相互作用。结果:成功构建了原核表达载体pET30a-LRP4胞内区和真核表达载体pEASY-Blunt M2-SNX17; 抗Myc单克隆抗体磁珠与His-LRP4混合后不发生共沉淀,如再与Myc-SNX17 混合则可发生共沉淀; 抗His单克隆抗体磁珠与Myc-SNX17混合后不发生共沉淀,再与His-LRP4胞内区混合可发生共沉淀。结论:原核表达的LRP4胞内区蛋白与真核表达的SNX17体外可发生直接结合。
Abstract:
Aim: To explore the interaction between LRP4 and SNX17 at the neuromuscular junction(NMJ)in patients with myasthenia gravis(MG).Methods: We constructed prokaryotic expression vector pET30a-LRP4 intracellular region,transformed it into E. coli BL21(DE3)bacteria and induced expression of His-LRP4 intracellular region with relative molecular weight of about 17 800; we constructed eukaryotic expression vector pEASY-Blunt M2-SNX17, transfected it into HEK293T cells and expressed Myc-SNX17 with relative molecular weight of about 53 000.Then we used two-way co-immunoprecipitation to verify whether there was interaction between LRP4 and SNX17.Results: We constructed prokaryotic expression vector pET30a-LRP4 intracellular region and eukaryotic expression vector pEASY-Blunt M2-SNX17 successfully; Anti-Myc-tag mAb-magnetic beads mixed with His-LRP4 did not occur co-precipitation, while those mixed with Myc-SNX17 showed co-precipitation; Anti-His-tag mAb-magnetic beads mixed with Myc-SNX17 did not occur co-precipitation, while those mixed with His-LRP4 intracellular region showed co-precipitation.Conclusion: Prokaryotic expression LRP4 intracellular region and eukaryotic expression SNX17 can combine directly in vitro.

参考文献/References:

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目 81471545; 河南省高等学校重点科研项目计划 16A320010,17A320046; 河南省科技攻关计划项目 172102 3010298; 河南省高校科技创新团队项目 16IRTSTHN022
#通信作者:高峰,男,1970年12月生,在读博士,研究员,研究方向:神经免疫病机制与临床,E-mail:gaoyuanshan@126.com; 王金兰,女,1968年9月生,博士,主任医师,研究方向:脑血管病及自身免疫性疾病,E-mail:wangjinlan08@126.com
更新日期/Last Update: 2017-03-20