Im Artikel <37A1D32E.517AB51A at rz.uni-potsdam.de>, Frank
=?iso-8859-1?Q?F=FCrst?= <ffrank at rz.uni-potsdam.de> schreibt:
>- coexpressing chaperones (a technology I am not an expert in, so
>somebody else should comment on that)
>That's just what I was about to suggest, because I have just finished reading a
recent CELL paper about the role of the DnaK chaperone in preventing protein
precipitation t very high local concentrations, as they occur at the
polyribosome... I admit that I am not an expert either.
>Or by denaturing and solubilizing your protein in the IBs and trying
>to renature it. If you're protein is of at most medium size and
>monomeric, I bet you can find conditions that allow refolding in
>sufficient yields.
Years ago, we were able to renature proteins within SDS-PAGE-Gels (!), up to
~100 kDa in size, by including BSA into the gel polymerization reaction (yes
indeed, most batches of BSA contain poorly characterized "chaperonoids", such
as prolyl isomerases and I-dunno-what-else) and soaking the gel for several
days in buffers containing some "alchemy", mostly low and decreasing
concentrations of mercaptoethanol. That was enough to regain lots of activity.
>I'm SignatureVirus 99! Copy me into your signature and join the fun!
HEY! Have you read Dawkins & Goodenough on "mind viruses", published a few
years ago in either NATURE or SCIENCE, I do not remember which? Read that and
have double fun! :-)
Good luck,
Rüdiger
===
Yours sincerely,
Chevalier Dr. Ruediger Marcus Flaig
University of Heidelberg
Email:
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"Tell truth and shame the devil." (Shakespeare, Henry IV.)
