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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.
NEDD8 is an important ubiquitinated protein in eukaryotes and is involved in the regulation of a series of important life processes in cells.Nowadays, the main method of obtaining NEDD8 is biocompatibility expression The disadvantage of the strategy is that the expression level is not high and the step of removing the extra purification tag is required.In this study, a homogeneous, high-affinity NEDD8 protein was obtained efficiently by combining the high-temperature assisted solid-phase synthesis with the one-pot ligation-desulfuration strategy based on the peptidic hydrazide linkage , Which provided a basis for further study of the structure and function of NEDD8-modified protein.