Serine promoted synthesis of peptide thioester-precursor on solid support for native chemical ligation.
Serine promoted synthesis of peptide thioester-precursor on solid support for native chemical ligation.
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DOI：
10.1039/c6sc02162j
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发表时间：
2017-01-01
期刊：
影响因子：
8.4
通讯作者：
Raj M
中科院分区：
文献类型：
--
作者：
Elashal HE;Sim YE;Raj M
Fmoc固相肽合成肽硫酯通过取代环氨基甲酸酯部分获得的C-末端丝氨酸的选择性活化。Fmoc固相肽合成硫酯对于通过天然化学连接进行蛋白质的化学合成是一个挑战。我们开发了一种利用Fmoc化学从固体载体直接合成肽硫酯的通用方法。肽硫酯合成通过主链酰胺链与丝氨酸侧基的选择性反应形成环状氨基甲酸酯部分来进行。活化的环状氨基甲酸酯部分经历硫醇置换以直接从固体载体产生硫酯。重要的是，该方法活化丝氨酸残基用于肽硫酯的合成;因此它是完全自动化的，并且不含使用Fmoc化学合成肽硫酯通常所需的树脂、接头、柄和非天然氨基酸的类型。所得硫酯不含差向异构化，并成功地应用于使用NCL合成较长的肽。
Fmoc solid phase peptide synthesis of peptide thioesters by displacement of the cyclic urethane moiety obtained by the selective activation of C-terminal serine. Fmoc solid phase peptide synthesis of thioesters for the chemical synthesis of proteins via native chemical ligation is a challenge. We have developed a versatile approach for direct synthesis of peptide thioesters from a solid support utilizing Fmoc chemistry. Peptide thioester synthesis is performed by the formation of a cyclic urethane moiety via a selective reaction of the backbone amide chain with the side group of serine. The activated cyclic urethane moiety undergoes displacement by a thiol to generate the thioester directly from the solid support. Importantly, the method activates the serine residue for the synthesis of peptide thioesters; thus it is fully automated and free of the types of resins, linkers, handles, and unnatural amino acids typically needed for the synthesis of peptide thioesters using Fmoc chemistry. The resulting thioester is free of epimerization and is successfully applied for the synthesis of longer peptides using NCL.