I would export the protein to the oxidative periplasm or alternatively oxidize the cys-cys-bond with Cu.
Best wishes
Kornelius
On Fri, 8 Feb 2008 10:44:21 -0500
Kendall Nettles <[log in to unmask]> wrote:
> I'm trying to engineer a disulfide bond into a protein that has several
> other cysteines.
>
> My question is whether there is a crystallization friendly reducing agent
> that can be used to prevent oxidation of the free cysteines without breaking
> the disulfide?
>
> Also, can I expect 100% disulfide formation from standard bacterial
> expression (assuming good geometry of the cysteines)?
>
>
> Thanks,
>
> Kendall
> --
> Kendall W. Nettles, PhD
> Assistant Professor
> Department of Cancer Biology
> The Scripps Research Institute
> 5353 Parkside Dr.
> Jupiter Fl 33458
----------------------------------------------
Kornelius Zeth
Max Planck Institute for Developmental Biology
Dept. Protein Evolution
Spemannstr. 35
72076 Tuebingen, Germany
[log in to unmask]
Tel -49 7071 601 323
Fax -49 7071 601 349
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