A new method for purification of functional recombinant GST-cyclophilin A protein from E. coli.
Indian J Biochem Biophys
;
2008 Dec; 45(6): 374-8
Article
in English
| IMSEAR
| ID: sea-27413
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.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Protein Binding
/
Rats
/
Recombinant Fusion Proteins
/
Cloning, Molecular
/
Protein Folding
/
Molecular Chaperones
/
Cyclophilin A
/
Escherichia coli
/
Formates
/
Glutathione Transferase
Language:
English
Journal:
Indian J Biochem Biophys
Year:
2008
Type:
Article
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