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

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...

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Autores principales: Mehlich, Alexander, Fang, Jie, Pelz, Benjamin, Li, Hongbin, Stigler, Johannes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750197/
https://www.ncbi.nlm.nih.gov/pubmed/33365300
http://dx.doi.org/10.3389/fchem.2020.587824
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author Mehlich, Alexander
Fang, Jie
Pelz, Benjamin
Li, Hongbin
Stigler, Johannes
author_facet Mehlich, Alexander
Fang, Jie
Pelz, Benjamin
Li, Hongbin
Stigler, Johannes
author_sort Mehlich, Alexander
collection PubMed
description 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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spelling pubmed-77501972020-12-22 Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein Mehlich, Alexander Fang, Jie Pelz, Benjamin Li, Hongbin Stigler, Johannes Front Chem Chemistry 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. Frontiers Media S.A. 2020-12-07 /pmc/articles/PMC7750197/ /pubmed/33365300 http://dx.doi.org/10.3389/fchem.2020.587824 Text en Copyright © 2020 Mehlich, Fang, Pelz, Li and Stigler. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Mehlich, Alexander
Fang, Jie
Pelz, Benjamin
Li, Hongbin
Stigler, Johannes
Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title_full Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title_fullStr Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title_full_unstemmed Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title_short Slow Transition Path Times Reveal a Complex Folding Barrier in a Designed Protein
title_sort slow transition path times reveal a complex folding barrier in a designed protein
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750197/
https://www.ncbi.nlm.nih.gov/pubmed/33365300
http://dx.doi.org/10.3389/fchem.2020.587824
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