Orsini G, Goldberg M E
J Biol Chem. 1978 May 25;253(10):3453-8.
The refolding and reoxidation of fully reduced and denatured chymotrypsinogen A have been studied in the presence of low concentrations of guanidine HCl or urea. Renaturation yields of 60 to 70% were observed when the reoxidation was facilitated by mixtures of reduced and oxidized glutathione. Refolding occurred within a narrow range of denaturant concentration (1.0 to 1.3 M guanidine HCl and 2 M urea) in which the native protein was shown to be stable, and the reduced protein was shown to regain the correct disulfide pairing. Renatured chymotrypsinogen is indistinguishable from the native zymogen in chromatographic behavior, potential chymotryptic activity, sedimentation coefficient, and spectral properties. The kinetics of renaturation were determined. Some of the protein species obtained at various times of renaturation were characterized as incorrectly oxidized molecules which could be renatured by thiol-catalyzed interchange of disulfide bonds.
在低浓度盐酸胍或尿素存在的情况下，对完全还原和变性的α-胰凝乳蛋白酶原A的复性和再氧化过程进行了研究。当由还原型和氧化型谷胱甘肽混合物促进再氧化时，观察到复性产率为60%至70%。在变性剂浓度的狭窄范围内（1.0至1.3 M盐酸胍和2 M尿素）发生复性，在此范围内天然蛋白质表现出稳定性，还原型蛋白质表现出重新获得正确的二硫键配对。复性后的α-胰凝乳蛋白酶原在色谱行为、潜在的胰凝乳蛋白酶活性、沉降系数和光谱性质方面与天然酶原无法区分。测定了复性动力学。在复性不同时间获得的一些蛋白质种类被表征为氧化错误的分子，它们可通过硫醇催化的二硫键交换而复性。
