Tbfmoc-蛋氨酸在泛素固相合成中的应用

J4 ›› 2011, Vol. 46 ›› Issue (7): 21-25.

Tbfmoc-蛋氨酸在泛素固相合成中的应用

金枝,胡乃峰,刘强,王朴*

山东大学药学院, 山东济南 250012

收稿日期:2010-12-25 出版日期:2011-07-20 发布日期:2011-09-08
通讯作者: 王朴(1955- ),男,教授,主要从事有机合成及多肽蛋白质合成研究. Email: puwang@sdu.edu.cn
作者简介:金枝(1986- )，女，硕士，主要从事有机合成及多肽合成. Email:jinzi-1986@163.com
基金资助:
国家自然科学基金资助项目(21072116);山东省自然科学基金资助项目(ZR2010BM040)

Application of TbfmocMetOH for solid phase synthesis of ubiquitin

JIN Zhi, HU Nai-feng, LIU Qiang, WANG Pu*

School of Pharmaceutical Science, Shandong University, Jinan 250012, Shandong, China

Received:2010-12-25 Online:2011-07-20 Published:2011-09-08

摘要/Abstract

摘要：

将合成的Tbfmoc蛋氨酸替代Fmoc蛋氨酸作为泛素的固相多肽化学合成原料。利用Tbfmoc泛素衍生物与各种杂质在凝胶过滤层析及紫外吸收的显著差异,通过凝胶过滤层析将Tbfmoc泛素从杂质中分离。在pH=85的温和条件下脱除 Tbfmoc 基团后,经氨基酸分析、质谱测定及高效液相色谱检测得到高纯度泛素产物。所用策略可有效克服采用 TbfmocCl时导致反应时间长、效率低、增加副产物及产物分离纯化难度大等弊病,为用途广泛的泛素化学合成及分离纯化提供了高效简便的新方法。

关键词: Tbfmoc蛋氨酸;泛素;固相多肽合成

Abstract:

The synthetic Tbfmoc-Met-OH is used to replace Fmoc-Met-OH as a building block for solid phase peptide synthesis of ubiquitin. Based on the significant differences in gel filtration and UV absorbance between the Tbfmocubiquitin and the impurities, Tbfmocubiquitin can be separated from the impurities using gel filtration easily and efficiently. After removing the Tbfmoc group under mild condition (pH=8.5), the ubiquitin product is obtained with high purity, which is confirmed by amino acid analysis, MS and HPLC measurements. The strategy used in the paper can overcome the Tbfmoc-Cl’s weak points, including long reaction time, low coupling efficiency, more by-product producing and product purification difficulty, and the established new method can be used for the synthesis, separation and purification of ubiquitin easily and efficiently.

Key words: Tbfmoc-Met-OH; ubiquitin; solid phase peptide synthesis

金枝,胡乃峰,刘强,王朴*. Tbfmoc-蛋氨酸在泛素固相合成中的应用[J]. J4, 2011, 46(7): 21-25.

JIN Zhi, HU Nai-feng, LIU Qiang, WANG Pu*. Application of TbfmocMetOH for solid phase synthesis of ubiquitin[J]. J4, 2011, 46(7): 21-25.

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