Synthesis of peptides and proteins without cysteine residues by native chemical ligation combined with desulfurization
Synthesis of peptides and proteins without cysteine residues by native chemical ligation combined with desulfurization
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DOI：
10.1021/ja003265m
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发表时间：
2001-01-31
影响因子：
15
通讯作者：
Dawson, PE
中科院分区：
文献类型：
--
作者：
Yan, LZ;Dawson, PE
带有 N 端 Cys 残基的未保护肽和 C 端硫酯之间的高度化学选择性反应能够实现复杂多肽的全合成和半合成。在这里，我们将这种天然化学连接方法的实用性扩展到不含半胱氨酸的肽。由于丙氨酸是蛋白质中常见的氨基酸，因此在该残基上进行连接将非常有用。为了实现这一目标，母体蛋白中的特定丙氨酸残基被半胱氨酸取代，以促进天然化学连接的合成。连接后，用 H-2/金属试剂对所得未保护的多肽产物进行选择性脱硫，将半胱氨酸残基转化为丙氨酸。这种方法提供了从半胱氨酰形式制备丙氨酰蛋白的通用方法，可用于化学合成多种多肽，如环状抗生素小菌素 J25、56 个氨基酸的链球菌蛋白 G B1 结构域和 110 个氨基酸核糖核酸酶变体、芽孢杆菌RNA酶的全化学合成所证明的那样。
The highly chemoselective reaction between unprotected peptides bearing an N-terminal Cys residue and a C-terminal thioester enables the total and semi-synthesis of complex polypeptides. Here we extend the utility of this native chemical ligation approach to non-cysteine containing peptides. Since alanine is a common amino acid in proteins, ligation at this residue would be of great utility. To achieve this goal, a specific alanine residue in the parent protein is replaced with cysteine to facilitate synthesis by native chemical ligation. Following ligation, selective desulfurization of the resulting unprotected polypeptide product with H-2/metal reagents converts the cysteine residue to alanine. This approach, which provides a general method to prepare alanyl proteins from their cysteinyl forms, can be used to chemically synthesize a variety of polypeptides, as demonstrated by the total chemical syntheses of the cyclic antibiotic microcin J25, the 56-amino acid streptococcal protein G B1 domain, and a variant of the 110-amino acid ribonuclease, barnase.