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 β-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.
作者
蒋环杰
王艳伟
陈家媛
胡丹
潘海
郭子龙
陈虎
Huanjie Jiang;Yanwei Wang;Jiayuan Chen;Dan Hu;Hai Pan;Zilong Guo;Hu Chen(Department of Physics,Wenzhou University,Wenzhou 325035,China;Center of Biomedical Physics,Wenzhou Institute,University of Chinese Academy of Sciences,Wenzhou 325000,China;Research Institute for Biomimetics and Soft Matter,Fujian Provincial Key Laboratory for Soft Functional Materials Research,Department of Physics,Xiamen University,Xiamen 361005,China)
基金
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)
research grant from Wenzhou Institute。
