Oxidative folding of cystine-rich peptides vs regioselective cysteine pairing strategies.
Oxidative folding of cystine-rich peptides vs regioselective cysteine pairing strategies.
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DOI：
10.1002/(sici)1097-0282(1996)40:2
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发表时间：
1996
期刊：
影响因子：
2.9
通讯作者：
L. Moroder;D. Besse;H. Musiol;S. Rudolph‐Böhner;F. Siedler
中科院分区：
文献类型：
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作者：
L. Moroder;D. Besse;H. Musiol;S. Rudolph‐Böhner;F. Siedler
合成多胱氨酸肽中的区域选择性半胱氨酸配对方法在过去几年中已取得进展，达到了允许至少三个特定的链间和链内二硫键桥接的效率。对各种多胱氨酸肽（如激素、蛋白酶抑制剂和毒素）的构象研究表明，这些由前体蛋白翻译后加工产生的生物活性肽被折叠成具有显着稳定性的小蛋白样致密球状结构。这有力地表明了这些肽家族的蛋白质结构域或子结构域特性，从而为在适当的实验条件下正确的氧化重折叠提供了足够的序列编码信息。通过对体内和体外蛋白质氧化重折叠机制和途径的深入研究，已经出现了模拟天然折叠再生的有效方法，不仅适用于完整蛋白质，还适用于蛋白质结构域和子结构域。事实上，在切除的蛋白质片段和相对低质量的翻译后加工产物的氧化折叠中获得的结果表明，如果根据氧化还原缓冲液、温度和能够破坏聚集体和稳定新生二级结构的添加剂来优化反应条件，则该过程确实是制备具有多个二硫键的肽的简单方法。此外，随着对稳定的小型天然胱氨酸框架的了解不断增加，使用它们代替人工模板将有助于具有特定构象和定制功能的合成微型蛋白的工程化。
The methodology of regioselective cysteine pairings in synthetic multiple-cystine peptides has progressed in the past years to an efficiency that allows for at least three specific inter- and intrachain disulfide bridgings. Conformational studies on various multiple-cystine peptides like hormones, protease inhibitors, and toxins revealed that these bioactive peptides, generated by posttranslational processing of precursor proteins, are folded into miniprotein-like compact globular structures of remarkable stability. This strongly suggests protein domain or subdomain properties of these families of peptides, and thus sufficient sequence-encoded information for correct oxidative refolding under appropriate experimental conditions. From intensive research on the mechanisms and pathways of oxidative refolding of proteins in vivo and in vitro, the efficient methods have emerged for simulating nature in the regeneration of native folds not only for intact proteins, but also for protein domains and subdomains. In fact, the results obtained in the oxidative folding of excised protein fragments and of relatively low mass products of posttranslational processings show that this procedure is indeed a simple way of preparing peptides with several disulfide bonds, if optimization of reaction conditions is performed in terms of redox buffer, temperature, and additives capable of disrupting aggregates and of stabilizing nascent secondary structures. Moreover, with increased knowledge about stable, small natural cystine frameworks, their use instead of artificial templates should facilitate engineering of synthetic miniproteins with specific conformation and tailored functions.