I know that conventional wisdom dictates that protease inhibitors should be added to lysis buffers when extracting protein to prevent sample degradation. That said, why are such inhibitors necessary when the lysis buffer is denaturing, considering that proteases are proteins? Shouldn't they also be denatured and incapable of proteolytic activitie?
For context, I am using a lysis buffer containing urea (8M), which is a denaturant.
A denatured protein is more susceptible to proteolysis than a protein in its properly folded state. If a protease is more stable to urea denaturation than the target protein, there could be some residual proteolysis, at least briefly.
I agree with Adam. In addition proteases are often remarkably stable compared to other (especially non-degradative) proteins. Anfinsen chose RNase A for some very good reasons!
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