Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating

Shun Hirota, Yukari Fujimoto, Jungkwon Choi, Naoki Baden, Noriko Katagiri, Masako Akiyama, Rinske Hulsker, Marcellus Ubbink, Toshihide Okajima, Teruhiro Takabe, Noriaki Funasaki, Yoshihito Watanabe, Masahide Terazima

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

A new method to investigate the initial protein folding dynamics is developed based on a pulsed laser light triggering method and a unique transient grating method. The side chain of the cysteine residue of apoplastocyanin (apoPC) was site-specifically modified with a 4,5-dimethoxy-2-nitrobenzyl derivative, where the CD and 2D NMR spectra showed that the modified apoPC was unfolded. The substituent was cleaved with a rate of about 400 ns by photoirradiation, which was monitored by the disappearance of the absorption band at 355 nm and the increase in the transient grating signal. After a sufficient time from the photocleavage reaction, the CD and NMR spectra showed that the native β-sheet structure was recovered. Protein folding dynamics was monitored in the time domain with the transient grating method from a viewpoint of the molecular volume change and the diffusion coefficient, both of which reflect the global structural change, including the protein-water interaction. The observed volume decrease of apoPC with a time scale of 270 μs is ascribed to the initial hydrophobic collapse. The increase in the diffusion coefficient (23 ms) is considered to indicate a change from an intermolecular to an intramolecular hydrogen bonding network. The initial folding process of apoPC is discussed based on these observations.

Original languageEnglish
Pages (from-to)7551-7558
Number of pages8
JournalJournal of the American Chemical Society
Volume128
Issue number23
DOIs
Publication statusPublished - 2006 Jun 14
Externally publishedYes

Fingerprint

Protein folding
Protein Folding
Nuclear magnetic resonance
Pulsed lasers
Absorption spectra
Hydrogen bonds
Hydrogen Bonding
Derivatives
Proteins
Cysteine
Lasers
Light
apoplastocyanin
Water

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating. / Hirota, Shun; Fujimoto, Yukari; Choi, Jungkwon; Baden, Naoki; Katagiri, Noriko; Akiyama, Masako; Hulsker, Rinske; Ubbink, Marcellus; Okajima, Toshihide; Takabe, Teruhiro; Funasaki, Noriaki; Watanabe, Yoshihito; Terazima, Masahide.

In: Journal of the American Chemical Society, Vol. 128, No. 23, 14.06.2006, p. 7551-7558.

Research output: Contribution to journalArticle

Hirota, S, Fujimoto, Y, Choi, J, Baden, N, Katagiri, N, Akiyama, M, Hulsker, R, Ubbink, M, Okajima, T, Takabe, T, Funasaki, N, Watanabe, Y & Terazima, M 2006, 'Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating', Journal of the American Chemical Society, vol. 128, no. 23, pp. 7551-7558. https://doi.org/10.1021/ja058788e
Hirota, Shun ; Fujimoto, Yukari ; Choi, Jungkwon ; Baden, Naoki ; Katagiri, Noriko ; Akiyama, Masako ; Hulsker, Rinske ; Ubbink, Marcellus ; Okajima, Toshihide ; Takabe, Teruhiro ; Funasaki, Noriaki ; Watanabe, Yoshihito ; Terazima, Masahide. / Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating. In: Journal of the American Chemical Society. 2006 ; Vol. 128, No. 23. pp. 7551-7558.
@article{691e8eafa7d747ffb47aae0b1fc06988,
title = "Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating",
abstract = "A new method to investigate the initial protein folding dynamics is developed based on a pulsed laser light triggering method and a unique transient grating method. The side chain of the cysteine residue of apoplastocyanin (apoPC) was site-specifically modified with a 4,5-dimethoxy-2-nitrobenzyl derivative, where the CD and 2D NMR spectra showed that the modified apoPC was unfolded. The substituent was cleaved with a rate of about 400 ns by photoirradiation, which was monitored by the disappearance of the absorption band at 355 nm and the increase in the transient grating signal. After a sufficient time from the photocleavage reaction, the CD and NMR spectra showed that the native β-sheet structure was recovered. Protein folding dynamics was monitored in the time domain with the transient grating method from a viewpoint of the molecular volume change and the diffusion coefficient, both of which reflect the global structural change, including the protein-water interaction. The observed volume decrease of apoPC with a time scale of 270 μs is ascribed to the initial hydrophobic collapse. The increase in the diffusion coefficient (23 ms) is considered to indicate a change from an intermolecular to an intramolecular hydrogen bonding network. The initial folding process of apoPC is discussed based on these observations.",
author = "Shun Hirota and Yukari Fujimoto and Jungkwon Choi and Naoki Baden and Noriko Katagiri and Masako Akiyama and Rinske Hulsker and Marcellus Ubbink and Toshihide Okajima and Teruhiro Takabe and Noriaki Funasaki and Yoshihito Watanabe and Masahide Terazima",
year = "2006",
month = "6",
day = "14",
doi = "10.1021/ja058788e",
language = "English",
volume = "128",
pages = "7551--7558",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "23",

}

TY - JOUR

T1 - Conformational changes during apoplastocyanin folding observed by photocleavable modification and transient grating

AU - Hirota, Shun

AU - Fujimoto, Yukari

AU - Choi, Jungkwon

AU - Baden, Naoki

AU - Katagiri, Noriko

AU - Akiyama, Masako

AU - Hulsker, Rinske

AU - Ubbink, Marcellus

AU - Okajima, Toshihide

AU - Takabe, Teruhiro

AU - Funasaki, Noriaki

AU - Watanabe, Yoshihito

AU - Terazima, Masahide

PY - 2006/6/14

Y1 - 2006/6/14

N2 - A new method to investigate the initial protein folding dynamics is developed based on a pulsed laser light triggering method and a unique transient grating method. The side chain of the cysteine residue of apoplastocyanin (apoPC) was site-specifically modified with a 4,5-dimethoxy-2-nitrobenzyl derivative, where the CD and 2D NMR spectra showed that the modified apoPC was unfolded. The substituent was cleaved with a rate of about 400 ns by photoirradiation, which was monitored by the disappearance of the absorption band at 355 nm and the increase in the transient grating signal. After a sufficient time from the photocleavage reaction, the CD and NMR spectra showed that the native β-sheet structure was recovered. Protein folding dynamics was monitored in the time domain with the transient grating method from a viewpoint of the molecular volume change and the diffusion coefficient, both of which reflect the global structural change, including the protein-water interaction. The observed volume decrease of apoPC with a time scale of 270 μs is ascribed to the initial hydrophobic collapse. The increase in the diffusion coefficient (23 ms) is considered to indicate a change from an intermolecular to an intramolecular hydrogen bonding network. The initial folding process of apoPC is discussed based on these observations.

AB - A new method to investigate the initial protein folding dynamics is developed based on a pulsed laser light triggering method and a unique transient grating method. The side chain of the cysteine residue of apoplastocyanin (apoPC) was site-specifically modified with a 4,5-dimethoxy-2-nitrobenzyl derivative, where the CD and 2D NMR spectra showed that the modified apoPC was unfolded. The substituent was cleaved with a rate of about 400 ns by photoirradiation, which was monitored by the disappearance of the absorption band at 355 nm and the increase in the transient grating signal. After a sufficient time from the photocleavage reaction, the CD and NMR spectra showed that the native β-sheet structure was recovered. Protein folding dynamics was monitored in the time domain with the transient grating method from a viewpoint of the molecular volume change and the diffusion coefficient, both of which reflect the global structural change, including the protein-water interaction. The observed volume decrease of apoPC with a time scale of 270 μs is ascribed to the initial hydrophobic collapse. The increase in the diffusion coefficient (23 ms) is considered to indicate a change from an intermolecular to an intramolecular hydrogen bonding network. The initial folding process of apoPC is discussed based on these observations.

UR - http://www.scopus.com/inward/record.url?scp=33745027642&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745027642&partnerID=8YFLogxK

U2 - 10.1021/ja058788e

DO - 10.1021/ja058788e

M3 - Article

VL - 128

SP - 7551

EP - 7558

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 23

ER -