Synthesis of a membrane protein with two transmembrane regions
Synthesis of a membrane protein with two transmembrane regions
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DOI：
10.1002/psc.381
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发表时间：
2002-04-01
影响因子：
2.1
通讯作者：
Aimoto, S
中科院分区：
文献类型：
--
作者：
Sato, T;Kawakami, T;Aimoto, S
采用硫酯法合成了具有两个跨膜结构域的膜蛋白。选择由 79 个氨基酸残基 (MW 8257) 组成的 F1F0 ATP 合酶亚基 c (Sub.c) 作为靶标。出于合成目的，使用 Boc 化学通过固相方法合成了两种结构单元 Boc-[Lys(34)(Boc)]-Sub.c(1-38)-SCH2CH2Co-Ala 和 Sub.c(39-79)。检查了跨膜肽的 RP-HPLC 纯化条件。结果发现，甲酸、1-丙醇和水的混合物与苯基柱的组合可用于分离跨膜肽。纯化的结构单元在氯化银、3,4-二氢3-氢-4-氧代-1,2,3-苯并三嗪（HOOBt）、N,N-二异丙基乙胺存在下在DMSO中缩合 给出产品，Sub.c。去除 Boc 基团后（产率 16%）。通过将反应温度提高至50℃，缩合反应的收率可提高至23％，当使用氯仿和甲醇的混合物作为溶剂时，缩合反应的收率可提高至26％。版权所有 (C) 2002 欧洲肽协会和 John Wiley Sons, Ltd.
A membrane protein with two transmembrane domains was synthesized by means of the thioester method. The F1F0 ATP synthase subunit c (Sub.c), which consists of 79 amino acid residues (MW 8257), was chosen as a target. For synthetic purposes, two building blocks, Boc-[Lys(34)(Boc)]-Sub.c(1-38)-SCH2CH2Co-Ala and Sub.c(39-79), were synthesized via solid-phase methods using Boc chemistry. RP-HPLC purification conditions for the transmembrane peptide were examined. As a result, a combination of a mixture of formic acid, 1-propanol and water with a phenyl column was found to be useful for Separating the transmembrane peptide, The purified building blocks were condensed in DMSO in the presence of silver chloride, 3,4-dihydro3-hydro-4-oxo-1,2,3-benzotriazine (HOOBt), N,N-diisopropylethylamine to give the product, Sub.c. after removal of Boc groups (yield 16%). The yield of the condensation reaction could be improved to 23% by raising the reaction temperature to 50degreesC, and to 26% when a mixture of chloroform and methanol was used as a solvent. Copyright (C) 2002 European Peptide Society and John Wiley Sons, Ltd.