y 08, 2008 10:44 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Engineering disulfide bonds
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
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 <[EMAIL PROTECTED]> wrote:
> I'm trying to engineer a disulfide bond into a protein that has several
> other cysteine
There are strains designed to provide a less-reducing (more
oxidizing) environment (Origami or any of it is "gami" derivatives
from Novagen).
They are deficient in the thioredoxin and glutathione reductases (I
think I recall...I didn't look it back up).
We've used them with good success f
On Fri, 2008-02-08 at 10:44 -0500, Kendall Nettles wrote:
> Also, can I expect 100% disulfide formation from standard bacterial
> expression (assuming good geometry of the cysteines)?
No, E. coli cells are a reducing environment.
-
=
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 format
