His-tagged proteins with less than six His

Andreas Savelsbergh asavelremove at gmx.de
Wed Oct 14 03:27:47 EST 1998


Hi Georg,

theoretically, "vicinal" Histidines are enough, so two to three should work.
With six you're just more on the safe side.... (since nobody really knows the
structure of the His-Tag??).

A colleague in my lab used to co-purify a certain E. coli protein with his
His-tagged protein, the other protein turned out to be more or less the only one
to have vicinal Histidines to exist in Eco.

Best regards,

Andreas

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






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