Citation: Xia Xu, Guiqian Yang, Zhen Zheng, Cody J. Wenthur, Jinyu Li, Gongyu Li. The sheet-to-helix transition is a potential gas-phase unfolding pathway for a multidomain protein CRM₁₉₇[J]. Chinese Chemical Letters, ;2025, 36(7): 110401. doi: 10.1016/j.cclet.2024.110401 shu

The sheet-to-helix transition is a potential gas-phase unfolding pathway for a multidomain protein CRM₁₉₇

Xia Xu ^{a, 1} ,
Guiqian Yang ^{b, 1} ,
Zhen Zheng ^c ,
Cody J. Wenthur ^d ,
Jinyu Li ^{b, *,} ,
Gongyu Li ^{a, *,}

a.
Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Center for Analytical Science, Frontiers Science Center for New Organic Matter, College of Chemistry, Nankai University, Tianjin 300071, China
b.
College of Chemistry, Fuzhou University, Fuzhou 350002, China
c.
School of Pharmacy, Tianjin Medical University, Tianjin 300070, China
d.
School of Pharmacy, University of Wisconsin-Madison, Wisconsin 53705, United States

j.li@fzu.edu.cn

ligongyu@nankai.edu.cn

Received Date: 30 April 2024
Revised Date: 5 August 2024
Accepted Date: 2 September 2024
Available Online: 3 September 2024

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Despite the expansive applications of gas-phase unfolding techniques, the molecular mechanism for the solvent-free forced unfolding pathway which substrate multidomain proteins usually adopt remains elusive at the secondary structure level. Herein, upon carefully selecting CRM₁₉₇ as a therapeutically-relevant model system containing multiple secondary structure-separated domains, we systematically examine its solvent-free unfolding pathway. Further-more, utilizing the hybrid of noncovalent chemical probing with niacinamide and ion mobility-mass spectrometry-guided all-atom molecular dynamics simulations, we map a nearly complete unfolding atlas for the conjugate vaccine carrier protein CRM₁₉₇ in a domain- and secondary structure-resolved manner. The totality of our data supports the preferential unfolding of the sheet-rich domain, indicating the dynamic transition from β-sheet to α-helix, and demonstrating that helix exhibit comparatively higher stability than β-sheets. We propose that this sheet-to-helix dynamic transition may be central to the gas-phase unfolding pathways of multidomain proteins, suggesting the need for systematic studies on additional multidomain protein systems.
- Sheet-to-helix transition,
- Ion mobility-mass spectrometry,
- Collision-induced unfolding,
- Molecular dynamics simulation,
- CRM₁₉₇
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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

The sheet-to-helix transition is a potential gas-phase unfolding pathway for a multidomain protein CRM197

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通讯作者: 陈斌, bchen63@163.com

The sheet-to-helix transition is a potential gas-phase unfolding pathway for a multidomain protein CRM₁₉₇