Int. J. Mol. Sci. 2013, 14(7), 13194-13212; doi:10.3390/ijms140713194
Article

Reinvestigation of the Oxidative Folding Pathways of Hen Egg White Lysozyme: Switching of the Major Pathways by Temperature Control

Department of Chemistry, School of Science, Tokai University, Kitakaname, Hiratsuka-shi, Kanagawa 259-1292, Japan
* Author to whom correspondence should be addressed.
Received: 2 May 2013; in revised form: 4 June 2013 / Accepted: 4 June 2013 / Published: 26 June 2013
(This article belongs to the Special Issue Protein Folding 2013)
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Abstract: It has been well established that in the oxidative folding of hen egg white lysozyme (HEL), which has four SS linkages in the native state (N), three des intermediates, i.e., des[76–94], des[64–80], and des [6–127], are populated at 20 °C and N is dominantly formed by the oxidation of des[64–80] and des[6–127]. To elucidate the temperature effects, the oxidative folding pathways of HEL were reinvestigated at 5–45 °C in the presence of 2 M urea at pH 8.0 by using a selenoxide reagent, DHSox. When reduced HEL was reacted with 1–4 equivalents of DHSox, 1S, 2S, 3S, and 4S intermediate ensembles with 1–4 SS linkages, respectively, were produced within 1 min. After the oxidation, 3S was slowly converted to the des intermediates with formation of the native structures through SS rearrangement. At 5 °C, des[76–94] was populated in the largest amount, but the oxidation to N was slower than that of des[64–80] and des[6–127]. At 35 °C, on the other hand, des[64–80] and des[6–127] were no longer stable, and only des[76–94] was populated. The results suggested that the major folding pathways of HEL can be switched from one to the other by temperature control.
Keywords: lysozyme; oxidative protein folding; temperature effect; selenoxide; reduction pulse; oxidation pulse

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MDPI and ACS Style

Arai, K.; Shibagaki, W.; Shinozaki, R.; Iwaoka, M. Reinvestigation of the Oxidative Folding Pathways of Hen Egg White Lysozyme: Switching of the Major Pathways by Temperature Control. Int. J. Mol. Sci. 2013, 14, 13194-13212.

AMA Style

Arai K, Shibagaki W, Shinozaki R, Iwaoka M. Reinvestigation of the Oxidative Folding Pathways of Hen Egg White Lysozyme: Switching of the Major Pathways by Temperature Control. International Journal of Molecular Sciences. 2013; 14(7):13194-13212.

Chicago/Turabian Style

Arai, Kenta; Shibagaki, Wataru; Shinozaki, Reina; Iwaoka, Michio. 2013. "Reinvestigation of the Oxidative Folding Pathways of Hen Egg White Lysozyme: Switching of the Major Pathways by Temperature Control." Int. J. Mol. Sci. 14, no. 7: 13194-13212.

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