I am using a GST-fusion construct for a metal-binding protein that we work on. We avoid a traditional 6x-His tag because Histidine residues have a fairly strong affinity for metal ions (hence, why Ni++ beads are able to pull out the tagged protein), and since we are analyzing a metal binding protein we do not want any interference from the Tag. However, our assays seem to indicate that our negative binding control (just GST alone) is binding small amounts of metal. Has anyone experienced anything like this or know of any literature that might delve into this issue a bit deeper? Thank you in advance.
Charles: GST has been known to bind metals, have you thought of using other fusion proteins/lectins such as maltose binding protein (MBP)?
Removing the GST tag would also solve your problem, if your construct provides a protease processing site.
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