STREP-Tag purification

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Wed Jun 26 21:03:48 EST 2002


Hi,

I'd rather not hawk our own work, but here's a structural paper -
Evdokimov, A. G., D. E. Anderson, K. M. Routzahn, and D. S. Waugh. 2001.
Structural basis for oligosaccharide recognition by Pyrococcus furiosus
maltodextrin-binding protein. J. Mol. Biol. 305: 891-904.

and here's some more stuff you might find useful -
Fox, J. D., R. B. Kapust, and D. S. Waugh. 2001. Single amino acid
substitutions on the surface of Escherichia coli maltose-binding protein can
have a profound impact on the solubility of fusion proteins. Protein Sci.
10: 622-630.

Acta Crystallogr D Biol Crystallogr 2002 Mar;58(Pt 3):392-7 Related
Articles, Books, LinkOut
Differential effects of short affinity tags on the crystallization of
Pyrococcus furiosus maltodextrin-binding protein.
Bucher MH, Evdokimov AG, Waugh DS.

I can send two of those to you as PDF's - the rest of the data isn't
published :)

Artem

"Philipp Wechner" <philipp.wechner at uibk.ac.at> wrote in message
news:3D19D2A0.340DE5F9 at uibk.ac.at...
>
>
> Artem Evdokimov schrieb:
>
> > > The DTT is not the biggest problem. I want to extract my protein from
> > inclusion
> > > bodies in E.coli. After treating the IB with 7M Urea or 5 M GuHCl i
tried
> > to
> > > get my protein by NTA-Chormatography (C-terminal 6xHis tag). However,
the
> > > protein does not bind too good - but its OK.
> > I understand your plight. The best you can do is try - there is no
telling
> > if STREP tag will work in your case, unless you dialyze into better
buffer.
> > have you considered fusing your protein to a thermophilic MBP, and
trying to
> > renature the fusion while keeping MBP alive ? PfuMBP does not really
> > denature in 5.5 M guanidinium hydrochloride at normal temperature, your
> > protein probably does...
> >
> > A.G.E.
>
> This sounds very interesting. I did an NCBI and google search on this mbp
out
> of pfu, but did not find anything. would be nice if you could tell me
where to
> find literature about this.
>
> thanks
>
>
>





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