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Re: [ccp4bb] Engineering disulfide bonds
 

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CCP4bb <-- 2008 <-- February 2008 <-- 08 February 2008
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Subject: Re: Engineering disulfide bonds
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
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Subject: Re: Engineering disulfide bonds
From: Kornelius Zeth kornelius {- dot -} zeth {- at -} TUEBINGEN {- dot -} MPG {- dot -} DE
Date: 2008-02-08
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 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
kornelius.zeth@tuebingen.mpg.de
Tel -49 7071 601 323
Fax -49 7071 601 349
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