Facile solid-phase synthesis of sulfated tyrosine-containing peptides: Part II. Total synthesis of human big gastrin-II and its C-terminal glycine-extended peptide (G34-Gly sulfate) by the solid-phase segment condensation approach.
Facile solid-phase synthesis of sulfated tyrosine-containing peptides: Part II. Total synthesis of human big gastrin-II and its C-terminal glycine-extended peptide (G34-Gly sulfate) by the solid-phase segment condensation approach.
复制标题
DOI：
10.1248/cpb.49.958
复制
发表时间：
2001-08
影响因子：
1.7
通讯作者：
K. Kitagawa;C. Aida;H. Fujiwara;T. Yagami;S. Futaki
中科院分区：
文献类型：
--
作者：
K. Kitagawa;C. Aida;H. Fujiwara;T. Yagami;S. Futaki
通过合成人大胃泌素-II（一种含34个残基的硫酸化酪氨酸[Tyr(SO3H)]肽），研究了基于芴9-基甲氧基羰基（Fmoc）的固相片段缩合方法在硫酸化肽制备中的应用。高度酸敏感的2-氯三苯甲基树脂（Clt树脂）专门用作锚定树脂，用于制备具有C端Pro残基的三个肽片段以及含有Tyr（SO3H）的树脂结合片段。通过使用PyBOP介导的偶联方案[PyBOP=苯并三唑氧基三(吡咯烷)鏻六氟磷酸盐，我们成功地缩合了每个片段并毫无困难地构建了34个残基的肽树脂。在低温下对完全保护的肽-树脂进行最终酸处理（90% TFA水溶液，0℃，8小时），可以将含有Tyr(SO3H)的肽从树脂上分离出来，并在硫酸盐劣化最小化的同时去除保护基团，得到粗硫酸化肽。经过一步 HPLC 纯化，可以从受保护的肽树脂中轻松获得高度均质的人大胃泌素-II，收率 14%。 C 端甘氨酸延伸胃泌素（G34-Gly 硫酸盐）的硫酸盐形式是胃泌素-II 的翻译后加工中间体，也通过片段缩合方法成功制备（产率 11%）。这些结果证明了片段缩合方案对于制备大的含 Tyr(SO3H) 的肽的有用性。
Application of the fluoren-9-ylmethoxycarbonyl (Fmoc)-based solid-phase segment condensation approach to the preparation of sulfated peptides was investigated through the synthesis of human big gastrin-II, a 34-residue sulfated tyrosine [Tyr(SO3H)]-containing peptide. Highly acid-sensitive 2-chlorotrityl resin (Clt resin) was exclusively employed as an anchor-resin for the preparation of the three peptide segments having the C-terminal Pro residue as well as of the Tyr(SO3H)-containing resin-bound segment. By using the PyBOP-mediated coupling protocol [PyBOP=benzotriazolyloxytris(pyrrolidino)phosphonium hexafluorophosphatel, we successively condensed each segment and constructed the 34-residue peptide-resin without any difficulty. The final acid treatment of the fully protected peptide-resin at low temperature (90% aqueous TFA, 0 degree C for 8 h), which can detach a Tyr(SO3H)-containing peptide from the resin and remove the protecting groups concurrently with minimum deterioration of the sulfate, afforded a crude sulfated peptide. After one-step HPLC purification, a highly homogeneous human big gastrin-II was easily obtained in 14% yield from the protected peptide-resin. The sulfate form of the C-terminal glycine-extended gastrin (G34-Gly sulfate), a posttranslational processing intermediate of gastrin-II, was also successfully prepared with the segment condensation approach (11% yield). These results demonstrated the usefulness of the segment condensation protocol for preparing large Tyr(SO3H)-containing peptides.