Oganesyan Natalia, Kim Sung-Hou, Kim Rosalind
Berkeley Structural Genomics Center, Physical Biosciences Division, Lawrence Berkeley National Laboratory, California 94720, USA.
J Struct Funct Genomics. 2005;6(2-3):177-82. doi: 10.1007/s10969-005-2827-3.
One major bottleneck in protein production in Escherichia coli for structural genomics projects is the formation of insoluble protein aggregates (inclusion bodies). The efficient refolding of proteins from inclusion bodies is becoming an important tool that can provide soluble native proteins for structural and functional studies. Here we report an on-column refolding method established at the Berkeley Structural Genomics Center (BSGC). Our method is a combination of an 'artificial chaperone-assisted refolding' method previously proposed and affinity chromatography to take advantage of a chromatographic step: less time-consuming, no filtration or concentration, with the additional benefit of protein purification. It can be easily automated and formatted for high-throughput process.
在用于结构基因组学项目的大肠杆菌蛋白质生产中，一个主要瓶颈是不溶性蛋白质聚集体（包涵体）的形成。从包涵体中有效重折叠蛋白质正成为一种重要工具，可为结构和功能研究提供可溶性天然蛋白质。在此，我们报告了在伯克利结构基因组学中心（BSGC）建立的一种柱上重折叠方法。我们的方法是将先前提出的“人工伴侣辅助重折叠”方法与亲和色谱相结合，以利用色谱步骤：耗时更少，无需过滤或浓缩，还具有蛋白质纯化的额外优势。它可以很容易地实现自动化并适用于高通量流程。
