Streamlined purification and characterization of Pyrococcus furiosus rubredoxins with different N-terminal modifications by reversed-phase HPLC
Streamlined purification and characterization of Pyrococcus furiosus rubredoxins with different N-terminal modifications by reversed-phase HPLC
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通过反相 HPLC 对具有不同 N 末端修饰的激烈火球菌红氧还蛋白进行简化纯化和表征
DOI：
10.1016/j.ab.2021.114128
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发表时间：
2021
影响因子：
2.9
通讯作者：
He Chunmao
中科院分区：
文献类型：
--
作者：
Lin Shaomin;He Chunmao
红毒杆菌毒素（Rds），像那些从毒杆菌（Pf），已被发现在大肠杆菌(E。大肠杆菌)作为不同n端形式的混合物，这可能会影响蛋白质的性质。典型的Rds纯化程序繁琐且收率低。本文提出了一种基于反相高效液相色谱（RP-HPLC）的简化纯化策略，该策略在预处理后只需一步纯化即可提供高收率和高分辨率。我们还表明，RP-HPLC可以作为一种有价值的工具来获得与pf - rd的热分解途径相关的信息。
Rubredoxins (Rds), like those fromPyrococcusfurious(Pf), have largely been found to be expressed inEscherichia coli(E. coli) as a mixture of differentN-terminal forms, which may affect the properties of the protein. The typical procedures for the purification of Rds are cumbersome and usually with low yield. We present herein a streamlined purification strategy based on the reversed-phase high performance liquid chromatography (RP-HPLC), which offers high yield and high resolution after simply one-step purification following pre-treatment. We also show that RP-HPLC can be a valuable tool to gain information related to the thermal decomposition pathway of Pf-Rds.