Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein

被引:13
作者
Mehlich, Alexander [1 ]
Fang, Jie [2 ]
Pelz, Benjamin [1 ]
Li, Hongbin [2 ]
Stigler, Johannes [3 ]
机构
[1] Tech Univ Munich, Phys Dept E22, Garching, Germany
[2] Univ British Columbia, Dept Chem, Vancouver, BC, Canada
[3] Ludwig Maximilians Univ Munchen, Gene Ctr Munich, Munich, Germany
来源
FRONTIERS IN CHEMISTRY | 2020年 / 8卷
关键词
protein folding; transition path analysis; artificial protein; transition state barrier; roughness; Kramers rate theory; ENERGY LANDSCAPE; FLUCTUATION THEOREM; SINGLE; FORCE; KINETICS; DIFFUSION; MODEL; TRAJECTORIES; FULL; DNA;
D O I
10.3389/fchem.2020.587824
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
De-novo designed proteins have received wide interest as potential platforms for nano-engineering and biomedicine. While much work is being done in the design of thermodynamically stable proteins, the folding process of artificially designed proteins is not well-studied. Here we used single-molecule force spectroscopy by optical tweezers to study the folding of ROSS, a de-novo designed 2x2 Rossmann fold. We measured a barrier crossing time in the millisecond range, much slower than what has been reported for other systems. While long transition times can be explained by barrier roughness or slow diffusion, we show that isotropic roughness cannot explain the measured transition path time distribution. Instead, this study shows that the slow barrier crossing of ROSS is caused by the population of three short-lived high-energy intermediates. In addition, we identify incomplete and off-pathway folding events with different barrier crossing dynamics. Our results hint at the presence of a complex transition barrier that may be a common feature of many artificially designed proteins.
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页数:11
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