removal of hexahistidine tag by peptidase

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Fri Feb 6 03:28:56 EST 1998


In article <34d88733.1031107 at newsserv.uni-bayreuth.de>,
Georg.Lipps at uni-bayreuth.de (Georg Lipps) wrote:

> Hi everybody,
> 
> does anyone has been successful in removing a C-terminal of N-terminal
> hexahistidine tag by limited exoproteolysis ?
> I have tried Aminopeptidase M and Carboxypeptidase A with my protein
> but have not been successful. Any hints are greatly appreciated !
> 
> Thanks in advance,
> 
> Georg

Hi Georg,
No idea about peptidases. But: I also had to remove a tag from my fusion
protein, and did that by constructing a derivative of my expression clone
with a factor Xa cleavage site (IEGR) right between the tag and the
insert. Simple cloning of two complementary oligos did the job (two days
of work only). Mail me if you are interested in a protocol.
Frank



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