Dirk Seegert wrote:
>> Hi there,
> I wonder wheather anybody could give me hints on how to get a monomeric
> recombinant protein into its natural occuring bioactive tetrameric form. Is
> it possible to solve this problem by production of tagged proteins? If so,
> which tags could be useful? Do I have to use specific cell lines?
> I would be happy for any advice.
You could try to fuse a tetrameric coiled coil to your subunits. Every
Subunit is fused to one helix, and of course you have to use a
modified sequence for the coiled-coil helix, so that it forms
tetramers instead of trimers.
Coiled-coil-sequences are usually derived from a protein called Gcmp4
(hope I didn't mix up letters), and if you need references, I could
ask a colleague who recently did this (but with the trimeric
original).
HTH,
Frank
--
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