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Protein Expr Purif. 2011 Feb;75(2):161-4. doi: 10.1016/j.pep.2010.09.006. Epub 2010 Sep 16.

Improved isolation of proteins tagged with glutathione S-transferase.

Author information

1
Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, USA.

Abstract

A common affinity tag used to express and purify fusion proteins is glutathione S-transferase. However, many researchers have reported difficulty eluting GST-tagged proteins from the affinity matrix. This report demonstrates that the problem likely is due to the propensity of glutathione S-transferase to dimerize combined with a propensity of the tagged protein to oligomerize, which results in formation of large oligomers of fusion protein that are chelated by the affinity matrix. The solution to the problem is to use S-butylglutathione instead of glutathione to elute, as S-butylglutathione binds more tightly to glutathione S-transferase and overcomes the chelate effect. Moreover, in contrast to glutathione, S-butylglutathione has no reducing capability that might inactivate a tagged protein.

PMID:
20849958
DOI:
10.1016/j.pep.2010.09.006
[Indexed for MEDLINE]

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