Thermal unfolding process of dihydrolipoamide dehydrogenase studied by fluorescence spectroscopy

Etsuko Nishimoto, Yoichi Aso, Toshiaki Koga, Shoji Yamashita

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The thermal unfolding pathway for dihydrolipoamide dehydrogenase (LipDH) isolated from Bacillus stearothermophilus was investigated focusing on the transient intermediate state characterized through time-resolved fluorescence studies. The decrease in ellipticity in the far UV region in the CD spectrum, the fluorescence spectral change of Trp-91 and FAD, and the thermal enzymatic inactivation curve consistently demonstrated that LipDH unfolded irreversibly on heat treatment at higher than 65°C. LipDH took a transient intermediate state during the thermal unfolding process which could refold back into the native state. In this state, the internal rotation of FAD was activated in the polypeptide cage and correspondingly LipDH showed a peculiar conformation. The transient intermediate state of LipDH characterized in time-resolved fluorescence depolarization studies showed very similar properties to the molten-globule state, which has been confirmed in many studies on protein folding.

Original languageEnglish
Pages (from-to)349-357
Number of pages9
JournalJournal of biochemistry
Volume140
Issue number3
DOIs
Publication statusPublished - Sep 1 2006

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Dihydrolipoamide Dehydrogenase
Fluorescence Spectrometry
Fluorescence spectroscopy
Flavin-Adenine Dinucleotide
Hot Temperature
Fluorescence
Protein folding
Depolarization
Bacilli
Geobacillus stearothermophilus
Conformations
Molten materials
Protein Folding
Heat treatment
Peptides

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Thermal unfolding process of dihydrolipoamide dehydrogenase studied by fluorescence spectroscopy. / Nishimoto, Etsuko; Aso, Yoichi; Koga, Toshiaki; Yamashita, Shoji.

In: Journal of biochemistry, Vol. 140, No. 3, 01.09.2006, p. 349-357.

Research output: Contribution to journalArticle

Nishimoto, Etsuko ; Aso, Yoichi ; Koga, Toshiaki ; Yamashita, Shoji. / Thermal unfolding process of dihydrolipoamide dehydrogenase studied by fluorescence spectroscopy. In: Journal of biochemistry. 2006 ; Vol. 140, No. 3. pp. 349-357.
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