Johnson Matthew, Geeves Michael A, Mulvihill Daniel P
School of Biosciences, University of Kent, Canterbury, UK.
Methods Mol Biol. 2013;981:193-200. doi: 10.1007/978-1-62703-305-3_15.
The majority of proteins in eukaryote cells are subjected to amino-terminal acetylation. This co-translational modification can affect the stability of a protein and also regulate its biological function. Amino-terminally acetylated recombinant proteins cannot be produced using prokaryote expression systems, such as E. coli, as these cells lack the appropriate N-α-terminal acetyltransferase complexes. Here we describe a simple protocol that allows the recombinant expression and purification of NatB-dependent amino-terminally acetylated proteins from E. coli.
真核细胞中的大多数蛋白质都要进行氨基末端乙酰化。这种共翻译修饰会影响蛋白质的稳定性,还能调节其生物学功能。由于原核生物表达系统(如大肠杆菌)缺乏合适的N-α-末端乙酰转移酶复合物,因此无法产生氨基末端乙酰化的重组蛋白。在此,我们描述了一种简单的方法,可用于从大肠杆菌中重组表达和纯化依赖于NatB的氨基末端乙酰化蛋白。