Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, Kumamoto 860-0082, Japan.
J Biochem. 2010 Mar;147(3):427-31. doi: 10.1093/jb/mvp184. Epub 2009 Nov 11.
Earlier, we formally established an effective refolding procedure for a protein by gradient removal of a solubilizer such as urea [Maeda et al. (1995) Effective renaturation of reduced lysozyme by gentle removal of urea. Protein Eng. 8, 201-205]. However, this procedure was less effective for unstable proteins. We developed here an excellent method to add protein stabilizer so as to get reasonable amounts of folded protein under the concentration of solubilizer where the unstable protein does not form aggregate. We examined many stabilizers and found that 60% of a concentrated (2.5 mg/ml) unstable protein can be refolded using 40% glycerol as the best stabilizer. This procedure can be widely applicable for the refolding of unstable proteins.
早些时候，我们通过梯度去除脲等溶剂[Maeda 等人，（1995）通过温和去除脲有效复性还原溶菌酶。蛋白质工程。8,201-205]正式建立了一种有效复性蛋白质的方法。然而，该方法对不稳定蛋白质的效果较差。我们在这里开发了一种极好的方法，可以添加蛋白质稳定剂，以便在不稳定蛋白质不会形成聚集体的溶剂浓度下获得合理量的折叠蛋白质。我们研究了许多稳定剂，发现 2.5mg/ml 浓度的不稳定蛋白质的 60%可以使用 40%的甘油作为最佳稳定剂进行复性。该程序可广泛适用于不稳定蛋白质的复性。
