A modified method for the purification of active large enzymes using the glutathione S-transferase expression system

作者：Deceglie Stefania; Lionetti Claudia; Roberti Marina; Cantatore Palmiro; Polosa Paola Loguercio^*

来源：Analytical Biochemistry, 2012, 421(2): 805-807.

DOI：10.1016/j.ab.2011.12.015

• GST fusion protein
• Protein affinity purification
• Mitochondrial RNA polymerase

The glutathione S-transferase (GST) fusion protein system is widely used for high-level expression and efficient purification of recombinant proteins from bacteria. However many GST-tagged proteins are insoluble, and the existing procedures, which employ a mixture of detergents to solubilize the molecules, frequently compromise their functional activity. A further limitation is that large proteins (%26gt;80 kDa) are poorly isolated by the current methods and are contaminated by truncated forms. To overcome these problems, we provide here an improved method for efficient purification of active large GST-tagged enzymes such as the 180-kDa GST-fused mitochondrial RNA polymerase.

• 出版日期2012-2-15