蛋白-DNA协同组装构建亚微米级复合结构

代江兵; 张丽霞; 毛秀海; 赵彦; 李柯; 谷沛霖; 郭琳洁; 李江; 钟超; 樊春海; 王丽华

doi:10.11777/j.issn1000-3304.2018.18251

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蛋白-DNA协同组装构建亚微米级复合结构

研究论文 | 更新时间：2021-01-26

- 蛋白-DNA协同组装构建亚微米级复合结构
- Submicrometre Superstructure Co-assembled from Protein and DNA
- 高分子学报 2019年50卷第4期页码：359-365
- 作者机构：
  
  1.中国科学院上海应用物理研究所中国科学院界面物理与技术重点实验室　上海 201800
  2.中国科学院大学　北京 100049
  3.上海嘉定区中心医院口腔科　上海 201800
  4.上海科技大学物质科学与技术学院　上海 201210
- 作者简介：
  
  E-mail: wanglihua@sinap.ac.cn Li-hua Wang, E-mail: wanglihua@sinap.ac.cn
- 基金信息：
  
  国家自然科学基金(基金号 61475181，U1532119，81471748)、国家重点研发计划(项目号 2016YFA0201200，2016YFA0400902)、中国科学院前沿科学重点研究(项目号 QYZDJ-SSW-SLH031-02)和中国科学院王宽诚率先人才计划-卢嘉锡国际团队项目资助()
- DOI：10.11777/j.issn1000-3304.2018.18251
  中图分类号：
- 纸质出版日期：2019-4，
  
  网络出版日期：2019-1-25，
  
  收稿日期：2018-11-23，
  
  修回日期：2018-12-28，
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代江兵, 张丽霞, 毛秀海, 赵彦, 李柯, 谷沛霖, 郭琳洁, 李江, 钟超, 樊春海, 王丽华. 蛋白-DNA协同组装构建亚微米级复合结构[J]. 高分子学报, 2019,50(4):359-365.

Jiang-bing Dai, Li-xia Zhang, Xiu-hai Mao, Yan Zhao, Ke Li, Pei-lin Gu, Lin-jie Guo, Jiang Li, Chao Zhong, Chun-hai Fan, Li-hua Wang. Submicrometre Superstructure Co-assembled from Protein and DNA[J]. Acta Polymerica Sinica, 2019,50(4):359-365.
代江兵, 张丽霞, 毛秀海, 赵彦, 李柯, 谷沛霖, 郭琳洁, 李江, 钟超, 樊春海, 王丽华. 蛋白-DNA协同组装构建亚微米级复合结构[J]. 高分子学报, 2019,50(4):359-365. DOI： 10.11777/j.issn1000-3304.2018.18251.

Jiang-bing Dai, Li-xia Zhang, Xiu-hai Mao, Yan Zhao, Ke Li, Pei-lin Gu, Lin-jie Guo, Jiang Li, Chao Zhong, Chun-hai Fan, Li-hua Wang. Submicrometre Superstructure Co-assembled from Protein and DNA[J]. Acta Polymerica Sinica, 2019,50(4):359-365. DOI： 10.11777/j.issn1000-3304.2018.18251.

摘要

以大肠杆菌菌毛蛋白CsgA组装形成的蛋白纤维为模板，引导不同数目的DNA四面体(tetrahedron DNA nanostructure，TDN)组装构建了蛋白-DNA亚微米复合结构. TDN经次氮基三乙酸(NTA)修饰后在Ni

的螯合作用下与CsgA蛋白单体结合，利用CsgA的自组装能力将TDN有序地排列在形成的蛋白纤维上. 原子力显微镜(atomic force microscopy，AFM)成像结果表明，控制TDN与CsgA的浓度比为1:500，可以得到单个TDN与蛋白纤维的组装产物. 将2个TDN通过杂交形成二聚体(dTDN)与CsgA蛋白进行组装，得到的亚微米复合结构保持了很好的直链形态，在蛋白纤维上连有3个dTDN结构的比例达44%.

Abstract

Functional supramolecular complexes co-assembled with multiple proteins and nucleic acids are ubiquitous in nature

such as ribosome and viruses. It is fundamentally important to understand and utilize these heterogeneous structures. Here

we reported a protein-DNA submicrometre superstructure fabricated from the self-assembly of protein CsgA and DNA nanostructures. By inspecting the physiological conditions of protein-DNA co-assembly

we found that the originally soluble CsgA could polymerize into insoluble fibers in a particular buffer (30 mmol/L Tris-HCl

450 mmol/L NaCl

pH = 7.2)

and such fibers benefited the storage of tetrahedron DNA nanostructure (TDN). Concentration and fibrillation time were optimized for the aggregation-free conversion of CsgA into amyloid fibers. Specifically

the optimum conversion of monomeric CsgA into micrometer-scale fibers could be realized at a concentration of 5 μmol/L. Meanwhile

atomic force microscopy (AFM) suggested that CsgA assembled into mature fibers in 5 days and formed larger aggregates after 7 days. The height of mature fibers and aggregates was about 3.9 and 6.1 nm

respectively. Afterwards

TDN was modified with NTA molecule and conjugated to CsgA through the chelation of Ni

His-tag

and NTA. A submicrometre complex CsgA fiber-dTDN was further generated by hybridizing two copies of TDN in dimeric structure (dTDN) through

-sheet interactions and Watson-Crick hybridization. This approach could fabricate a series of dTDN structures precisely without inducing the random aggregation of molecules. The yield of up to 44% was higher than that obtained from the direct connection of DNA modules

via

DNA technology. In summary

our findings demonstrated that CsgA fibers could act as a sort of novel scaffold for the assembly of protein-templated DNA nanostructure. Particularly

this model provides a deep insight into the generation of functional superstructures through self-assembly of protein and DNA-based building blocks.

关键词

协同组装DNA纳米技术CsgA蛋白蛋白-DNA复合结构

Keywords

Co-AssemblyDNA nanotechnologyCsgA proteinProtein-DNA complex

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