Laboratory of Organic Chemistry, ETH Zürich, Wolfgang Paulistrasse 10, CH-8006 Zürich, Switzerland.
J Biotechnol. 2010 Dec;150(4):481-9. doi: 10.1016/j.jbiotec.2010.09.956. Epub 2010 Oct 8.
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, pI 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 α-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.
小分子二硒化物在氧化蛋白质折叠中表现出巨大的潜力。为了探索其应用范围，使用含有二硒化物的氧化还原缓冲液来促进具有不同性质的蛋白质的折叠，这些性质包括大小、整体三级结构、二硫键数量、pI 值和体外折叠的难度。二硒化物能够催化所有测试蛋白质的氧化折叠，与类似的二硫键相比，显著提高了反应速率和产率。与谷胱甘肽的二硫键连接的二聚体（体外蛋白质折叠中最常用的氧化剂）相比，硒谷胱甘肽显著提高了生物技术重要蛋白质如水蛭素、溶菌酶、人表皮生长因子和干扰素 α-2a 的折叠效率。硒谷胱甘肽还增强了更具挑战性的靶标的复性，如牛血清白蛋白，其天然状态含有 17 个二硫键，以及抗体的 Fab 片段。在后一种情况下，毫摩尔浓度的硒谷胱甘肽能够与之前优化的氧化还原缓冲液提供的适度产量相匹配，该缓冲液含有毫摩尔浓度的谷胱甘肽。总之，这些不同蛋白质的折叠反应体现了二硒化物催化剂的优势和局限性。
