[1]孟兰霞,刘聪聪,张振涛.细菌CsgA蛋白促进α-Syn聚集[J].卒中与神经疾病杂志,2022,29(05):401-104,409.[doi:10.3969/j.issn.1007-0478.2022.05.001]
　Men Lanxia,Liu Congcong,Zhang Zhentao.Bacterial protein CsgA accelerates α-synuclein aggregation[J].Stroke and Nervous Diseases,2022,29(05):401-104,409.[doi:10.3969/j.issn.1007-0478.2022.05.001]
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细菌CsgA蛋白促进α-Syn聚集()
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《卒中与神经疾病》杂志[ISSN:1007-0478/CN:42-1402/R]

卷:
第29卷
期数:
2022年05期
页码:
401-104,409
栏目:
论著
出版日期:
2022-10-10

文章信息/Info

Title:
Bacterial protein CsgA accelerates α-synuclein aggregation
文章编号:
1007-0478(2022)05-0401-05
作者:
孟兰霞; 刘聪聪; 张振涛
430060 武汉大学人民医院神经内科［孟兰霞 刘聪聪 张振涛（通信作者）］
Author(s):
Men Lanxia;  Liu Congcong;  Zhang Zhentao
Departmental of Neurology, Renmin Hospital of Wuhan University, Wuhan 430060
关键词:
细菌; CsgA; R1结构域; 帕金森病; α-突触核蛋白
Keywords:
Bacteria; CsgA; R1 domain; Parkinson's disease; α-Synuclein
分类号:
R742.5
DOI:
10.3969/j.issn.1007-0478.2022.05.001
文献标志码:
A
摘要:
目的 探讨细菌CsgA蛋白对α-突触核蛋白（α-synulcien，α-Syn）聚集的影响及CsgA蛋白促进帕金森病发生的可能机制。 方法 在1.5 mg/mL的α-Syn单体蛋白中加入终浓度为0.01 mg/mL的CsgA蛋白R1结构域，采用硫黄素T染色法检测α-Syn的聚集；透射电镜分别观察R1，α-Syn和R1-α-Syn混合纤维的形态；分别将α-Syn纤维和R1-α-Syn混合纤维转导入稳定表达GFP标签的α-Syn的HEK293（Syn293）细胞中，观察对α-Syn聚集体形成的影响；分别用α-Syn纤维和R1-α-Syn混合纤维诱导Syn293细胞内形成聚集体，采用免疫荧光染色观察α-Syn聚集体是否具有路易小体的特征。 结果 R1蛋白可以明显促进α-Syn的聚集，并且这种聚集体具有路易小体的特征。 结论 细菌蛋白CsgA可促进α-Syn的聚集，这可能是细菌感染促进帕金森病发生的重要机制。
Abstract:
Objective To explore the effect of bacterial protein CsgA on the aggregation (α-Syn), and to illustrate the molecular mechanisms by which CsgA promote the onset of PD. Methods Experiment # 1: The aggregation of α-Syn (1.5 mg/ml) in the presence or absence of 0.01 mg/ml R1 as monitored by Thioflavin T fluorescence assay. Experiment # 2: The structure of the R1 pre-formed fibrils (PFFs), α-Syn PFFs, and R1-α-Syn mixed PFFs were observed by electron microscopy. Experiment # 3: α-Syn PFFs and R1-α-Syn mixed PFFs were transduced into HEK293 cells stably transfected with GFP-α-Syn (Syn293 cells). The formation of insoluble α-Syn aggregation was recorded. Experiment # 4: The characterization of intracellular aggregates induced by α-Syn PFFs and R1-α-Syn PFFs were investigated by immunofluorescence. Results R1 fragment of CsgA significantly promoted the aggregation of α-Syn. The R1-induced α-Syn PFFs show enhanced seeding activity in Syn293 cells. The aggregated indued by α-Syn PFFs and R1-α-Syn PFFs show characteristics of Lewy bodies. Conclusion R1, the key domain of CsgA, accelerates the aggregation of α-Syn. The CsgA-indued aggregation of α-Syn may play a role in the pathogenesis of Parkinson's disease.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金项目（81901090）
更新日期/Last Update: 2022-10-10