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
Artigo
em Inglês
| 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.
Texto completo:
DisponíveL
Índice:
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Assunto principal:
Ligação Proteica
/
Ratos
/
Proteínas Recombinantes de Fusão
/
Clonagem Molecular
/
Dobramento de Proteína
/
Chaperonas Moleculares
/
Ciclofilina A
/
Escherichia coli
/
Formiatos
/
Glutationa Transferase
Idioma:
Inglês
Revista:
Indian J Biochem Biophys
Ano de publicação:
2008
Tipo de documento:
Artigo
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