Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior

doi:10.1088/1674-1056/ad3dcd

中国物理B ›› 2024, Vol. 33 ›› Issue (7): 78201-078201.doi: 10.1088/1674-1056/ad3dcd

Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior

Huanjie Jiang(蒋环杰)^1,2, Yanwei Wang(王艳伟)¹, Jiayuan Chen(陈家媛)^1,2, Dan Hu(胡丹)^1,2, Hai Pan(潘海)², Zilong Guo(郭子龙)², and Hu Chen(陈虎)^2,3,†

1 Department of Physics, Wenzhou University, Wenzhou 325035, China;
2 Center of Biomedical Physics, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China;
3 Research Institute for Biomimetics and Soft Matter, Fujian Provincial Key Laboratory for Soft Functional Materials Research, Department of Physics, Xiamen University, Xiamen 361005, China

收稿日期:2024-01-25 修回日期:2024-03-18 接受日期:2024-04-12 出版日期:2024-06-18 发布日期:2024-06-18
通讯作者: Hu Chen E-mail:chenhu@xmu.edu.cn
基金资助:
This work was supported by the National Natural Science Foundation of China (Grant Nos. 12174322 to HC and 12204124 to ZG), 111 Project (Grant No. B16029), the Graduate Scientific Research Foundation of Wenzhou University (Grant No. 3162023003034 to JH), and research grant from Wenzhou Institute.

Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior

Huanjie Jiang(蒋环杰)^1,2, Yanwei Wang(王艳伟)¹, Jiayuan Chen(陈家媛)^1,2, Dan Hu(胡丹)^1,2, Hai Pan(潘海)², Zilong Guo(郭子龙)², and Hu Chen(陈虎)^2,3,†

1 Department of Physics, Wenzhou University, Wenzhou 325035, China;
2 Center of Biomedical Physics, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China;
3 Research Institute for Biomimetics and Soft Matter, Fujian Provincial Key Laboratory for Soft Functional Materials Research, Department of Physics, Xiamen University, Xiamen 361005, China

Received:2024-01-25 Revised:2024-03-18 Accepted:2024-04-12 Online:2024-06-18 Published:2024-06-18
Contact: Hu Chen E-mail:chenhu@xmu.edu.cn
Supported by:
This work was supported by the National Natural Science Foundation of China (Grant Nos. 12174322 to HC and 12204124 to ZG), 111 Project (Grant No. B16029), the Graduate Scientific Research Foundation of Wenzhou University (Grant No. 3162023003034 to JH), and research grant from Wenzhou Institute.

1. 2024-078201-SI.pdf(371KB)

摘要/Abstract

摘要： Single-molecule magnetic tweezers (MTs) have revealed multiple transition barriers along the unfolding pathway of several two-state proteins, such as GB1 and Csp. In this study, we utilized MTs to measure the force-dependent folding and unfolding rates of both protein L (PLWT) and its Y47W mutant (PLY47W) where the mutation point is not at the force-bearing $\beta$-strands. The measurements were conducted within a force range of 3-120 pN. Notably, the unfolding rates of both PLWT and PWY47W exhibit distinct force sensitivities below 50 pN and above 60 pN, implying a two-barrier free energy landscape. Both PLWT and PLY47W share the same force-dependent folding rate and the same transition barriers, but the unfolding rate of PLY47W is faster than that of PLWT. Our finding demonstrates that the residue outside of the force-bearing region will also affect the force-induced unfolding dynamics.

关键词: protein folding, magnetic tweezers, protein L

Abstract: Single-molecule magnetic tweezers (MTs) have revealed multiple transition barriers along the unfolding pathway of several two-state proteins, such as GB1 and Csp. In this study, we utilized MTs to measure the force-dependent folding and unfolding rates of both protein L (PLWT) and its Y47W mutant (PLY47W) where the mutation point is not at the force-bearing $\beta$-strands. The measurements were conducted within a force range of 3-120 pN. Notably, the unfolding rates of both PLWT and PWY47W exhibit distinct force sensitivities below 50 pN and above 60 pN, implying a two-barrier free energy landscape. Both PLWT and PLY47W share the same force-dependent folding rate and the same transition barriers, but the unfolding rate of PLY47W is faster than that of PLWT. Our finding demonstrates that the residue outside of the force-bearing region will also affect the force-induced unfolding dynamics.

Key words: protein folding, magnetic tweezers, protein L

中图分类号: (Single molecule manipulation of proteins and other biological molecules)

82.37.Rs

82.20.Db (Transition state theory and statistical theories of rate constants) 87.15.R- (Reactions and kinetics) 87.14.E- (Proteins)

Huanjie Jiang(蒋环杰), Yanwei Wang(王艳伟), Jiayuan Chen(陈家媛), Dan Hu(胡丹), Hai Pan(潘海), Zilong Guo(郭子龙), and Hu Chen(陈虎). Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior[J]. 中国物理B, 2024, 33(7): 78201-078201.

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Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior

Mutation in a non-force-bearing region of protein L influences force-dependent unfolding behavior

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