Protein crystallization with DNA templates--颗粒学报PARTICUOLOGY

Zhang, B., Yu, J., Pu, Z., Guo, W., Yang, R., Wen, Y., . . . Sun, H. (2024). Protein crystallization with DNA templates. Particuology, 86, 262-268. https://doi.org/10.1016/j.partic.2023.07.001

Protein crystallization with DNA templates

Bo Zhang ^a, Jia Yu ^a, Zhe Pu ^a, Wenhui Guo ^a, Ruoqi Yang ^a, Yu Wen ^a, Zhijiang Rong ^a, Hui Wang ^b, Xiaoli Wu ^a, Bo Wang ^b *, Hongyan Sun ^a *

a School of Environment and Resources, Taiyuan University of Science and Technology, Taiyuan, 030024, China
b School of Chemical Engineering and Technology, Taiyuan University of Science and Technology, Taiyuan, 030024, China

10.1016/j.partic.2023.07.001

Volume 86, March 2024, Pages 262-268

Received 23 March 2023, Revised 7 June 2023, Accepted 4 July 2023, Available online 14 July 2023, Version of Record 27 July 2023.

E-mail: wangbo@tyust.edu.cn; hongyansun@tyust.edu.cn

Highlights

• DNA polymeric additive templates effectively promotes protein crystallization efficiency.

• DNA promotes protein crystallization success rate, crystal number and mean size, crystallization rate and time.

• Better promotion of DNA templates when protein concentration is low.

• Facilitation: double-stranded DNA > single-stranded DNA, long-stranded DNA > short-stranded DNA.

Abstract

Protein crystallization plays a significant role in three-dimensional structural analysis and protein purification. It is important to increase the crystallization efficiency, which is possible by adding heterogeneous templates in crystallization systems. DNA is biologically compatible and artificially designable polymer, which is easy to extract. In this study, single- and double-stranded DNA of precise sequences were designed and used as templates to promote protein crystallization of lysozyme and catalase. Influence of DNA, single-stranded DNA with 10, 20, 40 bases and double-stranded DNA with 10, 20, 40 base pairs, were investigated. The success rate of obtaining crystals of lysozyme and catalase in equal period was significantly improved with the addition of DNA comparing without templates added. Double-stranded DNA led to higher nucleation rate than that with single-stranded DNA. The promotion of nucleation was more obvious at low concentration of protein solution and with longer chain DNA templates. Crystal number and crystallization rate was enhanced with addition of long double-stranded DNA templates. All the results confirm that DNA is an effective polymer additive to enhance protein crystallization, especially for the application of the scarce protein crystallization.

Graphical abstract

Keywords

DNA; Polymer; Additive; Protein; Crystallization

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