Citation: 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 shu

Submicrometre Superstructure Co-assembled from Protein and DNA

Jiang-bing Dai ^1,2,# ,
Li-xia Zhang ^3,# ,
Xiu-hai Mao ⁴ ,
Yan Zhao ¹ ,
Ke Li ⁴ ,
Pei-lin Gu ^1,2 ,
Lin-jie Guo ¹ ,
Jiang Li ¹ ,
Chao Zhong ⁴ ,
Chun-hai Fan ¹ ,
Li-hua Wang ^1,,

1.
CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800
2.
University of Chinese Academy of Sciences, Beijing 100049
3.
Jiading District Central Hospital, Shanghai 201800
4.
School of Physical Science and Technology, ShanghaiTech University, Shanghai 201210

Corresponding author: Li-hua Wang, wanglihua@sinap.ac.cn
Received Date: 23 November 2018
Revised Date: 28 December 2018
Available Online: 25 January 2019

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 Ni²⁺, His-tag, and NTA. A submicrometre complex CsgA fiber-dTDN was further generated by hybridizing two copies of TDN in dimeric structure (dTDN) through β-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.
- Co-Assembly,
- DNA nanotechnology,
- CsgA protein,
- Protein-DNA complex
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