Regulation of protein thermal stability and its potential application in the development of thermo-attenuated vaccines

Abstract

The coronavirus disease 2019 (COVID-19) pandemic has highlighted the importance of developing novel vaccines. An ideal vaccine should trigger an intense immune reaction without causing significant side effects. In this study we found that substitution of tryptophan located in the cores of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) protein structures with certain smaller amino acids resulted in variants with melting temperatures of 33-37 °C. An enzyme activity assay indicated that the proteolytic activity of the main proteinase (3CL^pro) decreased sharply when the environmental temperature exceeded the melting temperature, implying that other protein variants may lose most of their functions under the same conditions. This finding suggests that a virus variant containing engineered proteins with melting temperatures of 33-37 °C may only be functional in the upper respiratory tract where the temperature is about 33 °C, but will be unable to invade internal organs, which maintain temperatures above 37 °C, thus making it possible to construct temperature-sensitive attenuated vaccines.

Keywords

Protein melting temperature / Enzyme activity / SARS-CoV-2 N protein / 3CL^pro / Temperature sensitive attenuated vaccines

Data Availability Statement

All data generated or analyzed during this study are included in this published article.

Declaration of Competing Interest

The authors declare that they have no conflicts of interest with the contents of this article.

CRediT authorship contribution statement

Maofeng Wang: Writing - original draft, Visualization, Methodology, Investigation, Data curation. Cancan Wu: Writing - review & editing, Data curation. Nan Liu: Writing - review & editing. Xiaoqiong Jiang: Data curation. Hongjie Dong: Formal analysis. Shubao Zhao: Data curation. Chaonan Li: Investigation, Data curation. Sujuan Xu: Writing - review & editing. Lichuan Gu: Writing - review & editing, Supervision, Methodology, Funding acquisition, Conceptualization.

Acknowledgments

We are grateful for the funding from the Shandong Provincial Key Research and Development Program (2020CXGC011305).

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