Kasper Engkilde wrote:
>> Thank you for you interest...
>> I'm considering starting on a project where I will put a His-tag on my
> protein(s) but are conserned that I can't trust it to be active. My
> question is therefore:
>> What experience do you have with the activity of his-tagged proteins???
>> Good and bad are of cause equally welcome!
>> On before hand - Thank you for your time and help.
>> Sincerely Kasper Dreyer Engkilde
Depends on the enzyme and at which end you put the tag.
If your enzyme has the N- or C-terminus somehow stuck inside the main
protein-core, if one of the termini is involved in the active center or
necessary to uphold the structure or involved in the binding of other
subunits, than you are out of luck.
So far, I have been involved in two projects with two different classes
of enzymes, where the his-tag method was employed very successfully,
without any loss of activity.