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 language | English |
|---|---|
| Pages (from-to) | 349-357 |
| Number of pages | 9 |
| Journal | Journal of biochemistry |
| Volume | 140 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Sep 1 2006 |
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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 journal › Article
}
TY - JOUR
T1 - Thermal unfolding process of dihydrolipoamide dehydrogenase studied by fluorescence spectroscopy
AU - Nishimoto, Etsuko
AU - Aso, Yoichi
AU - Koga, Toshiaki
AU - Yamashita, Shoji
PY - 2006/9/1
Y1 - 2006/9/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=33750707565&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33750707565&partnerID=8YFLogxK
U2 - 10.1093/jb/mvj156
DO - 10.1093/jb/mvj156
M3 - Article
C2 - 16861247
AN - SCOPUS:33750707565
VL - 140
SP - 349
EP - 357
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 3
ER -