| Annexin A11 G38R及D40G突变通过降低蛋白稳定性引起肌萎缩侧索硬化症的发生 |
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| 引用本文： | 廖迪,廖巧,黄操,毕方方.Annexin A11 G38R及D40G突变通过降低蛋白稳定性引起肌萎缩侧索硬化症的发生[J].中南大学学报(医学版),2018,43(6):577-582. |
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| 作者姓名： | 廖迪 廖巧 黄操 毕方方 |
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| 作者单位： | 1. 中南大学湘雅医院神经内科，长沙 410008；2. 托马斯杰佛逊大学解剖学系，美国 宾夕法尼亚州 费城 19107 |
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| 基金项目： | 国家自然科学基金(81571256，81760238)。 |
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| 摘 要： | 目的：探讨Annexin A11(ANXA11) G38R及D40G突变引起肌萎缩侧索硬化症(amyotrophic lateral sclerosis，
ALS)的发病机制。方法：构建编码ANXA11 G38R，ANXA11 D40G突变蛋白及ANXA11野生型蛋白真核表达载体pANXA11-
G38R-Flag，pANXA11-D40G-Flag和pANXA11-Flag；将其转染至HEK293细胞后，用放线菌酮分别处理0，2，4，8 h；收
集蛋白采用Western印迹检测3种蛋白的含量变化情况，比较目标蛋白的半衰期；构建稳定表达ANXA11 G38R，ANXA11
D40G突变蛋白及ANXA11野生型蛋白的NSC-34细胞系，用Western印迹检测上述蛋白在NP-40裂解液中的溶解情况。结
果：ANXA11 G38R，ANXA11 D40G突变蛋白的半衰期较野生型蛋白缩短(P<0.05)，ANXA11 G38R蛋白和ANXA11 D40G
蛋白半衰期无明显差异(P>0.05)；ANXA11 G38R，ANXA11 D40G突变蛋白与ANXA11野生型蛋白在NP-40裂解液中均
未发现不可溶成分。结论：G38R和D40G突变降低了ANXA11蛋白的稳定性；G38R和D40G突变不影响ANXA11蛋白在
NP-40裂解液中的可溶性。
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| 关 键 词： | Annexin A11 肌萎缩侧索硬化症 半衰期 可溶性 |
Mutations of G38R and D40G cause amyotrophic lateral sclerosis by reducing Annexin A11 protein stability |
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| LIAO Di,LIAO Qiao,HUANG Cao,BI Fangfang.Mutations of G38R and D40G cause amyotrophic lateral sclerosis by reducing Annexin A11 protein stability[J].Journal of Central South University (Medical Sciences)Journal of Central South University (Medical Sciences),2018,43(6):577-582. |
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| Authors: | LIAO Di LIAO Qiao HUANG Cao BI Fangfang |
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| Institution: | 1. Department of Neurology, Xiangya Hospital, Central South University, Changsha 410008, China;
2. Department of Pathology, Anatomy & Cell Biochemistry, Th omas Jeff erson University, Philadelphia PA 19107, USA |
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| Abstract: | Objective: To explore the role of the mutations G38R and D40G of Annexin A11 (ANXA11) in
the onset of amyotrophic lateral sclerosis (ALS).
Methods: The plasmids expressing ANXA11 wild type protein, ANXA11 G38R protein and
ANXA11 D40G protein were constructed, respectively. The recombinant plasmids were thentransfected into HEK293 cells respectively followed by cycloheximide (CHX) treatment for 0, 2,
4 and 8 h. The protein expressions of ANXA11 wild type, ANXA11 G38R and ANXA11 D40G
mutations were determined by Western blot. Gray analysis by Image J was performed to compare
the half-life of each protein. The NSC-34 cell lines constantly expressing ANXA11 wild type
protein, ANXA11 G38R protein and ANXA11 D40G protein were established. The cells were
treated with NP-40 lysis buffer to examine the protein solubility by Western blot.
Results: Both ANXA11 G38R protein and ANXA11 D40G protein showed a shorter half-life
than ANXA11 wild type protein (P<0.05), while there was no difference between ANXA11 G38R
protein and ANXA11 D40G protein (P>0.05). There was no visible insoluble substance in the NP-
40 lysates for ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein.
Conclusion: G38R and D40G mutations reduce the stability of ANXA11 protein. G38R and
D40G mutations do not alter ANXA11 solubility. |
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| Keywords: | Annexin A11 amyotrophic lateral sclerosis half-life solubility |
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