Hmmm… is it really (physiological-like) binding, or your protein A is
aggregating/precipitating on the partner? Do you have a good negative control
(a similar protein to which protein A should not bind)? Also, as a general
rule, be careful about your detection method for the pull-down, don't
Dear All:
I have a general question about protein- protein interactions. I have two
proteins, A and B. A is a disordered protein while B is a well folded protein.
The binding between A and B has been approved by GST-pull down assay
previously. The strange thing is I cannot get them bind if
Dear Dee,
Some proteins with chaperone-like activity (perhaps your B?) can only
bind to partially folded proteins.
Probably A folds to a molten globule structure after 1-2 days. You can
check by spectroscopic techniques (ANS or Trp fluorescence, CD).
Hope that helps.
Cheers,
Clement
On
