protein folding - GST-fusion protein fluorescence (Aug/07/2006 )

Hi,
I will have to study the folding of a protein by fluorescence spectroscopy.
It is expressed as a GST-fusion protein. Should I cleave the GST tag before
performing the measurments (since gluthatione s-transferase contains Trp residues)?
Thanks

hi,

i think it's preferable for you to cleave off the gst tag since it is a reasonably big molecule (26 kDa, if i remember right). you will not be able to follow the folding of your protein in its native state if it has a gst tag on it, so you wont know if the pattern you see is really due to your protein going in to its native state or the new conformation it adopts due to the tag.

hi,

i think it's preferable for you to cleave off the gst tag since it is a reasonably big molecule (26 kDa, if i remember right). you will not be able to follow the folding of your protein in its native state if it has a gst tag on it, so you wont know if the pattern you see is really due to your protein going in to its native state or the new conformation it adopts due to the tag.