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.

参考文献

[1] Moore P B, Hendrickson W A, Henderson R and Brunger A T 2022 Science 375 507
[2] Bell G I 1978 Science 200 618
[3] Perl D, Welker C, Schindler T, Schröder K, Marahiel M A, Jaenicke R and Schmid F X 1998 Nat. Struct. Mol. Biol. 5 229
[4] Kim D E, Fisher C and Baker D 2000 J. Mol. Biol. 298 971
[5] McCallister E L, Alm E and Baker D 2000 Nat. Struct. Mol. Biol. 7 66973
[6] Guo Z, Hong H, Yuan G, Qian H, Li B, Cao Y, Wang W, Wu C and Chen H 2020 Phys. Rev. Lett. 125 198101
[7] Hong H, Guo Z, Sun H, Yu P, Su H, Ma X and Chen H 2021 Commun. Chem. 4 156
[8] Liu R, GarciaManyes S, Sarkar A, Badilla C L and Fernández J M 2009 Biophys. J. 96 3810
[9] Plaxco K W, Simons K T and Baker D 1998 J. Mol. Biol. 277 985
[10] Brockwell D J, Beddard G S, Paci E, West D K, Olmsted P D, Smith D A and Radford S E 2005 Biophys. J. 89 506
[11] Sun H, Guo Z, Hong H, Yu P, Xue Z and Chen H 2021 Biophys. Rep. 7 399
[12] Sun H, Guo Z, Hong H, Zhang Z, Zhang Y, Wang Y, Le S and Chen H 2023 Phys. Rev. Lett. 131 218402
[13] TapiaRojo R, Eckels E C and Fernández J M 2019 Proc. Natl. Acad. Sci. USA 116 7873
[14] Pierse C A and Dudko O K 2017 Phys. Rev. Lett. 118 088101
[15] Chen H, Fu H, Zhu X, Cong P, Nakamura F and Yan J 2011 Biophys. J. 100 517
[16] Ma X, Sun H, Hong H, Guo Z, Su H and Chen H 2022 Phys. Rev. E 106 024404
[17] Wikstroem M, Drakenberg T, Forsen S, Sjoebring U and Bjoerck L 1994 Biochemistry 33 14011
[18] O’Neill J W, Kim D E, Baker D and Zhang K Y 2001 Acta Crystallogr. D 57 480
[19] Scalley M L, Yi Q, Gu H, McCormack A, Yates J R and Baker D 1997 Biochemistry 36 3373
[20] Cao Y, Lam C, Wang M and Li H 2006 Angew. Chem. Int. Ed. 118 658
[21] Guo Z, Hong H, Sun H, Zhang X, Wu C, Li B, Cao Y and Chen H 2021 Nanoscale 13 11262
[22] Zakeri B, Fierer J O, Celik E, Chittock E C, SchwarzLinek U, Moy V T and Howarth M 2012 Proc. Natl. Acad. Sci. USA 109 E6907
[23] Chen H, Yuan G, Winardhi R S, Yao M, Popa I, Fernandez J M and Yan J 2015 J. Am. Chem. Soc. 137 3540
[24] Marko J F and Siggia E D 1995 Macromolecules 28 8759
[25] Best R B, Fowler S B, TocaHerrera J L and Clarke J 2002 Proc. Natl. Acad. Sci. USA 99 12143
[26] Lei H, He C, Hu C, Li J, Hu X, Hu X and Li H 2017 Angew. Chem. Int. Edit. 56 6117
[27] Cao Y, Kuske R and Li H 2008 Biophys. J. 95 782
[28] Schlierf M, Li H and Fernandez J M 2004 Proc. Natl. Acad. Sci. USA 101 7299

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

RichHTML

PDF (PC)

赞

补充材料

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 11

Metrics

本文评价

推荐阅读 0

[1]	Lujun Zou(邹禄军), Jiajun Lu(陆伽俊), and Xiulian Xu(徐秀莲). Effect of chaperone-client interaction strength on Hsp70-mediated protein folding[J]. 中国物理B, 2023, 32(11): 118701-118701.
[2]	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[J]. 中国物理B, 2021, 30(7): 78201-078201.
[3]	张晓峰, 郭子龙, 余平, 李秋实, 周昕, 陈虎. Different potential of mean force of two-state protein GB1 and downhill protein gpW revealed by molecular dynamics simulation[J]. 中国物理B, 2020, 29(7): 78701-078701.
[4]	毛雯雯, 吕丽花, 季永运, 李有泉. Quantum intelligence on protein folding pathways[J]. 中国物理B, 2020, 29(1): 18702-018702.
[5]	卢禹锟, 周昕, 欧阳钟灿. Smoothing potential energy surface of proteins by hybrid coarse grained approach[J]. 中国物理B, 2017, 26(5): 50202-050202.
[6]	冉诗勇, 贾俊丽. A multi-field approach to DNA condensation[J]. 中国物理B, 2015, 24(12): 128702-128702.
[7]	李伟, Wong Wei-Juan, Lim Ci-Ji, 车海鹏, 李明, 严洁, 王鹏业. Catch-bond behavior of DNA condensate under tension[J]. 中国物理B, 2015, 24(12): 128704-128704.
[8]	郑伟谋. Proteins：From sequence to structure[J]. 中国物理B, 2014, 23(7): 78705-078705.
[9]	刘景发. Structure optimization by heuristic algorithm in a coarse-grained off-lattice model[J]. 中国物理B, 2009, 18(6): 2615-2621.
[10]	江凡, 李南. Protein structural codes and nucleation sites for protein folding[J]. 中国物理B, 2007, 16(2): 392-404.
[11]	张庆利, 王莉, 翁羽翔, 邱祥冈, 王渭池, 阎吉祥. Nanosecond-time-resolved infrared spectroscopic study of fast relaxation kinetics of protein folding by means of laser-induced temperature-jump[J]. 中国物理B, 2005, 14(12): 2484-2490.