"Vladimir Grubor @optushome.com.au>" <forwards<nospam> wrote in message
news:a8r30j$o3$1 at ariel.ucs.unimelb.edu.au...
> I heard from experienced people that it is best to slowly decrease the
> denatureation gradient, rather than do a sudden transition.
For some proteins it might be crucial to decrease denaturant concentration
rapidly instead. Protein folding in nature is generally a rapid process that
takes only milliseconds to complete. The problem in correct refolding is
that if conditions are non-optimal, one or more misfolded conformations may
get sequestered from the folding process by forming stable aggregates. That
is why it might be necessary to do refolding in the presence of chemicals
(e.g. detergents) that prevent aggregation. I believe that in most cases
when slow dyalysis worked, it was simply because there were no or very small
number of misfolded protein conformations that form aggregates.
Emir
