Effect of DTT on protein structure/function independent of disulfide bond reduction

Kartik Chandran kartik_chandran at hms.harvard.edu
Mon Mar 11 19:23:23 EST 2002


Hi there,

I'm investigating the effect of an engineered disulfide bond on the
hemolytic activity of the protein I'm studying. In order to reduce the
buried disulfide, I'm having to treat with 50 mM DTT at 37šC for 15-20 min
prior to addition of red blood cells (to test hemolytic potential of the
protein). 

However, I get no activity in the presence of DTT even with a control
protein that lacks the two engineered cysteine residues. There are no other
disulfide bonds in the protein.

I'm wondering whether DTT may have effects on my protein independent of
reduction of the disulfide bond. If anyone has any ideas or input into my
problem, that would be great!

Thanks!

Kartik chandran

kartik_chandran at hms.harvard.edu


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