摘要/Abstract
二硫键的氧化折叠是合成二硫键构象锁定多肽的关键步骤. 前人发展的二硫键氧化折叠策略主要有一次氧化折叠、多次氧化折叠和一锅法氧化折叠. 目前对三种策略复性效率和收率等的比较性研究较少. 分别采用三种氧化折叠策略制备目标蜈蚣毒素多肽RhTx. 结果表明, 两次氧化折叠策略的分离收率高于一次和一锅法氧化折叠策略, 一锅法氧化折叠策略可能会导致较大比例的错误折叠. 探索了数十毫克量级RhTx的高效制备方法, 为进一步探索RhTx靶向TRPV1的结构机制等研究提供了工具分子. 此外, 对三种氧化折叠策略进行了系统比较, 为二硫键构象锁定多肽的合成提供了参考.
关键词: 二硫键, 二硫键构象锁定多肽, 氧化折叠, RhTx, 固相合成, 巯基保护基
The critical step for the synthesis of disulfide-containing peptides is the efficient construction of one or multiple disulfide bridges. Generally, the folding of disulfide bonds could be achieved by three chemical strategies,i.e. one-step oxidative folding strategy, multi-step oxidative folding strategy, and one-pot oxidative folding strategy. Because few comparative studies have been conducted on efficiencies of three strategies, the systematical research is desirable. Three folding strategies were separately applied for the preparation of centipede toxin RhTx. The results showed that the isolated yield of two-step oxidative folding strategy was higher than those of one-step and one-pot oxidative folding strategies. Besides, the one-pot oxidative folding strategy may induce severe misfoldings. The circular dichroism (CD) and activity tests indicated that disulfide bonds are critical for the structure and activity of RhTx. In addition, the efficient preparation of RhTx on tens of milligrams scale was achieved, affording molecular tools for the further biological and biophysical studies of RhTx targeting TRPV1. Overall, three mainstream oxidative folding strategies were systematically studied, which provided a valuable reference for the synthesis of disulfide-containing peptides.
Key words: disulfide bond, disulfide-containing peptide, oxidative folding, RhTx, solid phase peptide synthesis, thiol protecting group
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