On Wed, 8 Dec 1999, Alex Blais wrote:
> Hi,
>> I am trying to purify a recombinant protein fused to GST overproduced in
> E.coli. I have some problems during the purification steps, and I
> hypothesised that homodimers formation mediated by the GST moiety could
> be the cause of the problem.
>
I can't help you with the dimer problem, but the formation of dimer by
GST has been shown to affect the folding of protein.
See
Murray et al (95) PNAS, 92, 7337-7341
A few useful related articles:
Murray et al (98) Nature structure biology 5, 778-782
Hayes et al (99) J mol. biol. 285 1857-1867
> Does anybody know what conditions could be used to minimize the
> formation of such dimers ?
>> I already know that there are 4 cysteines in the gst monomer and I know
> that they are not involved in dimer formation, since mutation to serines
> still allows dimer formation (these are results published by other
> groups). I also know that in 2 M urea, the dimers are still allowed
> (this is my own experience).
>> Thanks a lot for any suggestions
>> Alexandre Blais
> Graduate Student
> Centre de Recherche du CHUL
> Québec, Canada
>>>
