Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale

Abstract In order to directly observe the refolding kinetics from a partially misfolded state to a native state in the bottom of the protein-folding funnel, we used a “caging” strategy to trap the β-sheet structure of ubiquitin in a misfolded conformation. We used molecular dynamics simulation to ge...

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Autores principales:	Eric H.-L. Chen, Tony T.-Y. Lu, Jack C.-C. Hsu, Yufeng Jane Tseng, T.-S. Lim, Rita P.-Y. Chen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7e49e2049803490d8d197737a1428dac
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spelling	oai:doaj.org-article:7e49e2049803490d8d197737a1428dac2021-12-02T15:06:10ZDirectly monitor protein rearrangement on a nanosecond-to-millisecond time-scale10.1038/s41598-017-08385-02045-2322https://doaj.org/article/7e49e2049803490d8d197737a1428dac2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08385-0https://doaj.org/toc/2045-2322Abstract In order to directly observe the refolding kinetics from a partially misfolded state to a native state in the bottom of the protein-folding funnel, we used a “caging” strategy to trap the β-sheet structure of ubiquitin in a misfolded conformation. We used molecular dynamics simulation to generate the cage-induced, misfolded structure and compared the structure of the misfolded ubiquitin with native ubiquitin. Using laser flash irradiation, the cage can be cleaved from the misfolded structure within one nanosecond, and we monitored the refolding kinetics of ubiquitin from this misfolded state to the native state by photoacoustic calorimetry and photothermal beam deflection techniques on nanosecond to millisecond timescales. Our results showed two refolding events in this refolding process. The fast event is shorter than 20 ns and corresponds to the instant collapse of ubiquitin upon cage release initiated by laser irradiation. The slow event is ~60 μs, derived from a structural rearrangement in β-sheet refolding. The event lasts 10 times longer than the timescale of β-hairpin formation for short peptides as monitored by temperature jump, suggesting that rearrangement of a β-sheet structure from a misfolded state to its native state requires more time than ab initio folding of a β-sheet.Eric H.-L. ChenTony T.-Y. LuJack C.-C. HsuYufeng Jane TsengT.-S. LimRita P.-Y. ChenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eric H.-L. Chen Tony T.-Y. Lu Jack C.-C. Hsu Yufeng Jane Tseng T.-S. Lim Rita P.-Y. Chen Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
description	Abstract In order to directly observe the refolding kinetics from a partially misfolded state to a native state in the bottom of the protein-folding funnel, we used a “caging” strategy to trap the β-sheet structure of ubiquitin in a misfolded conformation. We used molecular dynamics simulation to generate the cage-induced, misfolded structure and compared the structure of the misfolded ubiquitin with native ubiquitin. Using laser flash irradiation, the cage can be cleaved from the misfolded structure within one nanosecond, and we monitored the refolding kinetics of ubiquitin from this misfolded state to the native state by photoacoustic calorimetry and photothermal beam deflection techniques on nanosecond to millisecond timescales. Our results showed two refolding events in this refolding process. The fast event is shorter than 20 ns and corresponds to the instant collapse of ubiquitin upon cage release initiated by laser irradiation. The slow event is ~60 μs, derived from a structural rearrangement in β-sheet refolding. The event lasts 10 times longer than the timescale of β-hairpin formation for short peptides as monitored by temperature jump, suggesting that rearrangement of a β-sheet structure from a misfolded state to its native state requires more time than ab initio folding of a β-sheet.
format	article
author	Eric H.-L. Chen Tony T.-Y. Lu Jack C.-C. Hsu Yufeng Jane Tseng T.-S. Lim Rita P.-Y. Chen
author_facet	Eric H.-L. Chen Tony T.-Y. Lu Jack C.-C. Hsu Yufeng Jane Tseng T.-S. Lim Rita P.-Y. Chen
author_sort	Eric H.-L. Chen
title	Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
title_short	Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
title_full	Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
title_fullStr	Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
title_full_unstemmed	Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
title_sort	directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/7e49e2049803490d8d197737a1428dac
work_keys_str_mv	AT erichlchen directlymonitorproteinrearrangementonananosecondtomillisecondtimescale AT tonytylu directlymonitorproteinrearrangementonananosecondtomillisecondtimescale AT jackcchsu directlymonitorproteinrearrangementonananosecondtomillisecondtimescale AT yufengjanetseng directlymonitorproteinrearrangementonananosecondtomillisecondtimescale AT tslim directlymonitorproteinrearrangementonananosecondtomillisecondtimescale AT ritapychen directlymonitorproteinrearrangementonananosecondtomillisecondtimescale
_version_	1718388591645687808

Directly monitor protein rearrangement on a nanosecond-to-millisecond time-scale

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