STREP-Tag purification

Philipp Wechner philipp.wechner at uibk.ac.at
Thu Jun 27 04:24:46 EST 2002


Thanks for the literature. I am unable to download the PDF from the journals
page. Could you send me a copy to my email adress? philipp.wechner at uibk.ac.at

thanks a lot


Artem Evdokimov schrieb:

> 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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