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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

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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 2015)
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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 lysozyme; oxidative protein folding; temperature effect; selenoxide; reduction pulse; oxidation pulse
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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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