Diselenides as universal oxidative folding catalysts of diverse proteins
Diselenides as universal oxidative folding catalysts of diverse proteins
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DOI：
10.1016/j.jbiotec.2010.09.956
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发表时间：
2010-12-01
影响因子：
4.1
通讯作者：
Hilvert, Donald
中科院分区：
文献类型：
--
作者：
Beld, Joris;Woycechowsky, Kenneth J.;Hilvert, Donald
小分子二硒醚作为蛋白质氧化折叠的催化剂显示出相当大的潜力。为了探索它们的范围，使用含二硒化物的氧化还原缓冲液来促进性质不同的蛋白质的折叠，所述性质例如大小、总体三级结构、二硫键的数目、pl值和体外折叠的难度。二硒醚能够催化所有测试的蛋白质的氧化折叠，相对于类似的二硫化物，在速率和产率方面都提供显著增加。与谷胱甘肽的二硫键连接的二聚体（体外蛋白质折叠最常用的氧化剂）相比，硒代谷胱甘肽在生物技术上重要的蛋白质（如水蛭素、溶菌酶、人表皮生长因子和干扰素α-2a）的折叠中提供了显著提高的效率。硒代葡萄糖酸还可增强更具挑战性的靶标的复性，例如天然状态含有17个二硫键的牛血清白蛋白和抗体的Fab片段。在后一种情况下，微摩尔量的硒代谷胱甘肽能够匹配由先前优化的氧化还原缓冲液提供的适度产率，所述氧化还原缓冲液含有毫摩尔水平的谷胱甘肽。总之，这些不同蛋白质的折叠反应显示了二硒化物催化剂的优点和局限性。(C)2010爱思唯尔有限公司版权所有。
Small-molecule diselenides show considerable potential as catalysts of oxidative protein folding. To explore their scope, diselenide-containing redox buffers were used to promote the folding of proteins that varied in properties such as size, overall tertiary structure, number of disulfide bonds, pl value, and difficulty of in vitro folding. Diselenides are able to catalyze the oxidative folding of all proteins tested, providing significant increases in both rate and yield relative to analogous disulfides. Compared to the disulfide-linked dimer of glutathione (the most commonly used oxidant for in vitro protein folding), selenoglutathione provided markedly improved efficiencies in the folding of biotechnologically important proteins such as hirudin, lysozyme, human epidermal growth factor and interferon alpha-2a. Selenoglutathione also enhances the renaturation of more challenging targets such as bovine serum albumin, whose native state contains 17 disulfide bonds, and the Fab fragment of an antibody. In the latter case, micromolar amounts of selenoglutathione are able to match the modest yield provided by a previously optimized redox buffer, which contains millimolar levels of glutathione. Taken together, the folding reactions of these diverse proteins exemplify the advantages and limitations of diselenide catalysts. (C) 2010 Elsevier B.V. All rights reserved.