Chemical synthesis of proteins in solution.
Chemical synthesis of proteins in solution.
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DOI：
10.1002/(sici)1097-0282(1999)51:4
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发表时间：
1999
期刊：
影响因子：
2.9
通讯作者：
S. Sakakibara
中科院分区：
文献类型：
--
作者：
S. Sakakibara
描述了一种适用于蛋白质溶液合成的新策略的开发，其中整个分子由大小范围为约10个残基的完全保护的片段组装而成。每个片段被设计为具有Boc-肽-OPac（Pac：苯甲酰甲基）的共同结构，并且所有侧链官能团都被基于Bzl的基团保护。描述了用于溶解完全保护的链段的新型溶剂体系，其中在溶液中的链段缩合反应可以顺利进行。除去Boc或Pac基团后，使用1-乙基-3-（3’-二甲基氨基丙基）-碳二亚胺/3，4-二氢-3-羟基-4-氧代-1，2，3-苯并三嗪方法将片段偶联在一起以获得整个序列。然后通过HF除去侧链保护基团，并使释放的肽进行折叠反应以获得天然构象。应用该策略成功合成了123个氨基酸残基的人血管生成素、121个氨基酸残基的人中期因子、136个氨基酸残基的人多效生长因子和238个氨基酸残基的水母绿色荧光蛋白。
Development of a novel strategy suitable for the solution synthesis of proteins is described, wherein the entire molecule is assembled from fully protected segments in the size range of about 10 residues. Each segment is designed so as to have a common structure of Boc-peptide-OPac (Pac: phenacyl) and all of the side-chain functional groups are protected by Bzl-based groups. New types of solvent systems are described for dissolving fully protected segments in which the segment condensation reactions in solution can be carried out smoothly. After removal of the Boc or Pac group, the segments are coupled together to obtain the entire sequence using the 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide/3, 4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine method. The side-chain protecting groups are then removed by HF and the liberated peptide is subjected to folding reactions to obtain the native conformation. Applying the strategy, the 123-residue human angiogenin, the 121-residue human midkine, the 136-residue human pleiotrophin, and the 238-residue Aequoria green fluorescent protein were synthesized successfully.