多肽酰肼法合成翻译后修饰物蛋白nedd8 - 上海有机化学研究所.pdf

DOI: 10.6023/cjoc201605013 研究简报 有机化学 Chinese Journal of Organic Chemistry NOTE 多肽酰肼法合成翻译后修饰物蛋白NEDD8 管超建 王 涛 王 君* 李宜明* (合肥工业大学生物与医学工程学院 合肥 230009) 摘要 NEDD8(神经前体细胞表达发育下调蛋白 8)是真核生物体内一种重要的类泛素化蛋白, 参与调控细胞内一系列 重要的生命过程. 目前获得 NEDD8 的主要方法是生物重组表达, 该策略的缺点在于表达量不高且需要除去外加纯化 标签的步骤. 本研究结合高温辅助固相合成技术与基于多肽酰肼连接的一锅连接-脱硫策略, 较为高效地获得了均一性 的NEDD8 蛋白, 为进一步研究 NEDD8 修饰蛋白的结构与功能提供了基础. 关键词 NEDD8; 高温辅助固相合成; 多肽酰肼连接; 一锅连接脱硫 Synthesis of Post-translational Modifier Protein NEDD8 via Ligation of Peptide Hydrazides Guan, Chaojian Wang, Tao Wang, Jun* Li, Yiming* (Department of Pharmacy , School of Biological and Medical Engineering, Hefei University of Technology, Hefei 230009) Abstract As an important ubiquitin-like modifier protein in eukaryotic organisms, NEDD8 (neural precursor cell expressed developmentally down-regulated 8) is involved in regulating a series of important life processes in cells. Nowadays, the major method for obtaining NEDD8 is recombinant protein expression. However, the yield is relatively low and the recombined tag for purification needs to be removed in an extra step. In present work, NEDD8 protein was first synthesized in homogeneity by using high temperature assisted solid-phase peptide synthesis (SPPS) combining with the one-pot ligation-desulfurization strategy. This method lays the foundation for the study of NEDD8 modified proteins in future. Keywords NEDD8; high temperature assisted SPPS; ligation of peptide hydrazide; one-pot ligation-desulfurizat