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A new method for purification of functional recombinant GST-cyclophilin A protein from E. coli

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Title A new method for purification of functional recombinant GST-cyclophilin A protein from E. coli
 
Creator Lee, Seonmin
Han, Xuezhe
Choi, Kyu Jin
Ding, Yan
Choi, Taegyu
Tak, Eunyoung
Lee, Jisun
Ha, Joohun
Kim, Sung Soo
Lee, Jinhwa
 
Subject Chaperone activity
Cyclophilin A
Formic acid
Gluthatione-S-transferase fusion protein
PPIase activity
 
Description 374-378
The expression of glutathione-S-transferase (GST) fusion protein is extensively utilized in the study of protein-protein interactions. In the commonly used purification method, the overexpressed GST fusion protein is bound to the glutathione (GSH)-coupled resins via affinity chromatography, and then eluted by an excessive quantity of reduced GSH. However, this technique has certain limitations, such as low product purity, retention of GSH in the sample, as well as relatively high cost. To overcome these limitations, in this study, elution buffer containing 2% formic acid was utilized rather than GSH to elute the GST-fusion protein, and thereafter the acidic samples were neutralized using collecting buffer. By using this method, highly purified GST-cyclophilin A (CypA) fusion protein was obtained, without affecting the structural and functional characteristics such as PPIase and chaperone activities. Moreover, the procedure is also cost-effective, due to the low cost of formic acid as compared with GSH.
 
Date 2008-12-31T05:45:27Z
2008-12-31T05:45:27Z
2008-12
 
Type Article
 
Identifier 0301-1208
http://hdl.handle.net/123456789/2703
 
Language en_US
 
Publisher CSIR
 
Source IJBB Vol.45(6) [December 2008]