His-tagged proteins with less than six His

[Per Mygind]

Georg Wille wrote:

> Dear Bionetters,
>
> I'm about to attach an His-tag to the N-Terminus of a protein. Has anyone
> ever heard of using _less_ than six residues successfully. I would like to
> use as few as possible in order not to distort the enzymatic properties of
> the protein too much and shifting its pI as little as possible.
>
> The protein is a dimer and its N-termini are fairly close together, so maybe
> three His at each N-Terminus are maybe almost as good as six.
>
> Any experience? I would appreciate suggestions! Or maybe a reference to read
> about this method.
>
> Thanks a lot!
>
>      Georg

I have no reference for this, but I can remember, that only two histidines are
needed for nondenaturing purification
using Ni-chelate material. Contact me if you need a reference. Bear in mind,
that the affinity decreases, so that you might get an increase in contaminating
proteins.

Good luck

Per Mygind

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If you are are not part of the solution, you are part of the precipitate
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Per Mygind, Cand.scient

Department of Medical Microbiology and Immunology
The Bartholin Building, University of Aarhus, Denmark
phone : 89 42 17 47, fax   : 86 19 61 28

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