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Title: Enzymatic biosynthesis and immobilization of polyprotein verified at the single-molecule level
Authors: Deng, Yibing
Wu, Tao
Wang, Mengdi
Shi, Shengchao
Yuan, Guodong
Li, Xi
Chong, Hanchung
Wu, Bin
Zheng, Peng
Keywords: Atomic Force Microscopy
Science::Biological sciences
Issue Date: 2019
Source: Deng, Y., Wu, T., Wang, M., Shi, S., Yuan, G., Li, X., . . . Zheng, P. (2019). Enzymatic biosynthesis and immobilization of polyprotein verified at the single-molecule level. Nature Communications, 10(1), 2775-. doi:10.1038/s41467-019-10696-x
Series/Report no.: Nature Communications
Abstract: The recent development of chemical and bio-conjugation techniques allows for the engineering of various protein polymers. However, most of the polymerization process is difficult to control. To meet this challenge, we develop an enzymatic procedure to build polyprotein using the combination of a strict protein ligase OaAEP1 (Oldenlandia affinis asparaginyl endopeptidases 1) and a protease TEV (tobacco etch virus). We firstly demonstrate the use of OaAEP1-alone to build a sequence-uncontrolled ubiquitin polyprotein and covalently immobilize the coupled protein on the surface. Then, we construct a poly-metalloprotein, rubredoxin, from the purified monomer. Lastly, we show the feasibility of synthesizing protein polymers with rationally-controlled sequences by the synergy of the ligase and protease, which are verified by protein unfolding using atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS). Thus, this study provides a strategy for polyprotein engineering and immobilization.
DOI: 10.1038/s41467-019-10696-x
Rights: © 2019 The Author(s). Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
Fulltext Permission: open
Fulltext Availability: With Fulltext
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