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IJBB Vol.45(6) [December 2008] >

Title: A new method for purification of functional recombinant GST-cyclophilin A protein from E. coli
Authors: Lee, Seonmin
Han, Xuezhe
Choi, Kyu Jin
Ding, Yan
Choi, Taegyu
Tak, Eunyoung
Lee, Jisun
Ha, Joohun
Kim, Sung Soo
Lee, Jinhwa
Keywords: Chaperone activity
Cyclophilin A
Formic acid
Gluthatione-S-transferase fusion protein
PPIase activity
Issue Date: Dec-2008
Publisher: CSIR
Abstract: 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.
Page(s): 374-378
ISSN: 0301-1208
Source:IJBB Vol.45(6) [December 2008]

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