Structural characteristic of folding/unfolding intermediate of pokeweed anti-viral protein revealed by time-resolved fluorescence

Shuzo Matsumoto, Yuka Taniguchi, Yukihiro Fukunaga, Hiromichi Nakashima, Keiichi Watanabe, Shoji Yamashita, Etsuko Nishimoto

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The structural feature of unfolding intermediate of pokeweed anti-viral protein (PAP) was characterized using time-resolved fluorescence spectroscopic methods to elucidate protein folding/unfolding process. CD and fluorescence spectra consistently demonstrated that the unfolding of PAP completed at 4 M of guanidine hydrochloride (GuHCl). The fluorescence resonance energy transfer (FRET) and time-resolve fluorescence depolarization analysis of Trp208 and Trp237 located in the C-terminal domain of PAP suggested that peculiar unfolding intermediate populated before reaching to the unfolding state. The FRET distance of Trp237 to Tyr182 was extended to more than 28 Å with keeping the compact conformation in the unfolding intermediate state populated in the presence of 2 M GuHCl. On the other hand, Trp208 maintained the energy transfer pair with Tyr72 near the active site, although the rotational freedom was increased a little. There results suggest that the most distinguished structural feature of the unfolding intermediate of PAP is the separation of C-terminal domain from N-terminal domain. FRET and fluorescence depolarization studies also showed that C-terminal domain would be more separated to liberate the segmental motions of Trp208 and Trp237 distinctly at the unfolding state.

Original languageEnglish
Pages (from-to)407-415
Number of pages9
JournalJournal of Fluorescence
Volume23
Issue number3
DOIs
Publication statusPublished - May 1 2013

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Fluorescence Resonance Energy Transfer
Fluorescence
energy
Guanidine
Depolarization
Protein folding
Protein Unfolding
CD
Energy Transfer
Protein Folding
Energy transfer
Conformations
Catalytic Domain
pokeweed antiviral protein
time

All Science Journal Classification (ASJC) codes

  • Spectroscopy
  • Biochemistry
  • Clinical Biochemistry
  • Medicine(all)

Cite this

Structural characteristic of folding/unfolding intermediate of pokeweed anti-viral protein revealed by time-resolved fluorescence. / Matsumoto, Shuzo; Taniguchi, Yuka; Fukunaga, Yukihiro; Nakashima, Hiromichi; Watanabe, Keiichi; Yamashita, Shoji; Nishimoto, Etsuko.

In: Journal of Fluorescence, Vol. 23, No. 3, 01.05.2013, p. 407-415.

Research output: Contribution to journalArticle

Matsumoto, Shuzo ; Taniguchi, Yuka ; Fukunaga, Yukihiro ; Nakashima, Hiromichi ; Watanabe, Keiichi ; Yamashita, Shoji ; Nishimoto, Etsuko. / Structural characteristic of folding/unfolding intermediate of pokeweed anti-viral protein revealed by time-resolved fluorescence. In: Journal of Fluorescence. 2013 ; Vol. 23, No. 3. pp. 407-415.
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AU - Watanabe, Keiichi

AU - Yamashita, Shoji

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