Folding mechanism of canine milk lysozyme studied by circular dichroism and fluorescence spectroscopy

Masaharu Nakao, Munehito Arai, Takumi Koshiba, Katsutoshi Nitta, Kunihiro Kuwajima

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We have studied the guanidine hydrochloride-induced equilibrium unfolding and the kinetics of refolding of canine milk lysozyme by circular dichroism and fluorescence spectroscopy. The thermodynamic analysis of the equilibrium unfolding measured by circular dichroism and fluorescence has shown that unfolding is represented by a three-state mechanism and that the intermediate state of canine milk lysozyme is remarkably more stable than the intermediates observed in other lysozyme and α-lactalbumin. In the kinetic refolding of this protein, there are at least two kinetic intermediates; a burst-phase intermediate accumulated within the dead time (4 ms) of the measurement and an intermediate that has been observed during the kinetics with a rate constant of 10-20 s-1 after the burst phase. This result is apparently in contrast with those previously observed in the kinetic refolding of α-lactalbumin and equine lysozyme that show only the burst-phase intermediate. The relationship between the extraordinarily stable equilibrium molten globule and the kinetic folding intermediates will be discussed.

Original languageEnglish
Pages (from-to)183-193
Number of pages11
JournalSpectroscopy
Volume17
Issue number2-3
DOIs
Publication statusPublished - Jan 1 2003

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Circular dichroism spectroscopy
Fluorescence spectroscopy
Muramidase
Kinetics
Lactalbumin
Guanidine
Dichroism
Milk
Molten materials
Rate constants
Fluorescence
Thermodynamics

All Science Journal Classification (ASJC) codes

  • Spectroscopy

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Folding mechanism of canine milk lysozyme studied by circular dichroism and fluorescence spectroscopy. / Nakao, Masaharu; Arai, Munehito; Koshiba, Takumi; Nitta, Katsutoshi; Kuwajima, Kunihiro.

In: Spectroscopy, Vol. 17, No. 2-3, 01.01.2003, p. 183-193.

Research output: Contribution to journalArticle

Nakao, Masaharu ; Arai, Munehito ; Koshiba, Takumi ; Nitta, Katsutoshi ; Kuwajima, Kunihiro. / Folding mechanism of canine milk lysozyme studied by circular dichroism and fluorescence spectroscopy. In: Spectroscopy. 2003 ; Vol. 17, No. 2-3. pp. 183-193.
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