Renaturation of Lysozyme with a Protein Disulfide Isomerase Chaperone Results in Enzyme Super Activity

Takezawa, Aya; Ohshima, Yuji; Sudo, Tomoya; Asami, Osamu; Nohara, Daisuke

doi:10.1263/jbb.106.503

Cited by 1 publication

(1 citation statement)

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“…The experimental procedure previously described [7,33] was followed with slight modification. To a solution of HEL (12 mg) dissolved in 0.7 mL of a 100 mM Tris–HCl/1 mM EDTA buffer solution at pH 8.0 containing 8 M urea as a denaturant was added an excess amount of DTT red (12 mg).…”

Section: Methodsmentioning

confidence: 99%

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

Arai

Shibagaki

Shinozaki

et al. 2013

IJMS

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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.

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Section: Methodsmentioning

confidence: 99%