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Equilibrium folding and unfolding dynamics to reveal detailed free energy landscape of src SH3 protein by magnetic tweezers |
| Huanhuan Su(苏环环)1,†, Hao Sun(孙皓)1,†, Haiyan Hong(洪海燕)1, Zilong Guo(郭子龙)2,3, Ping Yu(余平)1, and Hu Chen(陈虎)1,2,3,‡ |
1 Institute for Biomimetics and Soft Matter, Fujian Provincial Key Laboratory for Soft Functional Materials Research, Department of Physics, Xiamen University, Xiamen 361005, China;
2 Center of Biomedical Physics, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China;
3 Oujiang Laboratory, Wenzhou 325000, China |
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Abstract Src SH3 protein domain is a typical two-state protein which has been confirmed by research of denaturant-induced unfolding dynamics. Force spectroscopy experiments by optical tweezers and atomic force microscopy have measured the force-dependent unfolding rates with different kinds of pulling geometry. However, the equilibrium folding and unfolding dynamics at constant forces has not been reported. Here, using stable magnetic tweezers, we performed equilibrium folding and unfolding dynamic measurement and force-jump measurement of src SH3 domain with tethering points at its N- and C-termini. From the obtained force-dependent transition rates, a detailed two-state free energy landscape of src SH3 protein is constructed with quantitative information of folding free energy, transition state barrier height and position, which exemplifies the capability of magnetic tweezers to study protein folding and unfolding dynamics.
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Received: 17 March 2021
Revised: 15 April 2021
Accepted manuscript online: 26 April 2021
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PACS:
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82.37.Rs
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(Single molecule manipulation of proteins and other biological molecules)
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82.20.Db
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(Transition state theory and statistical theories of rate constants)
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87.15.R-
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(Reactions and kinetics)
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87.14.E-
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(Proteins)
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| Fund: Project supported by the National Natural Science Foundation of China (Grant Nos. 11874309 and 11474237) and the 111 Project (Grant No. B16029). |
Corresponding Authors:
Hu Chen
E-mail: chenhu@xmu.edu.cn
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Cite this article:
Huanhuan Su(苏环环), Hao Sun(孙皓), Haiyan Hong(洪海燕), Zilong Guo(郭子龙), Ping Yu(余平), and Hu Chen(陈虎) Equilibrium folding and unfolding dynamics to reveal detailed free energy landscape of src SH3 protein by magnetic tweezers 2021 Chin. Phys. B 30 078201
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