remove his-tag

J. Arturo García-Horsman arturo.garcia-horsman at uku.fi
Wed May 2 06:57:12 EST 2001


Hi Slava,

Several companies now a days sell their his-tagged expression vectors with a
protease recognition site sequence engineered right before (or after, depending if
it is 3' or 5' his-tagging) the his tag sequence, so you can cleave the tail with
an specific protease. Of course you have to test first if this kind of tagging
won't affect folding/activity of your protein, or if it does, if cleaving would
reactivate it. Also, problems can arise if your protein also has internal sites for
this protease, but you can always make it resistant (again, if this does not
affecting folding/activity). I don't think you can eliminate the his tag otherwise,
but I might be wrong. Limited proteolysis experiments are very difficult to set up,
using an exo-peptidases, and I think they are difficult to reproduce. Although, if
you are lucky to have a protein which has already a natural specific protease site
near the end and that the cleavage product would be active, then you're all set.

Arturo

_______________________________________________________________

Arturo Garcia-Horsman, Ph.D.
Research Specialist
Department of Pharmacology and Toxicology
University of Kuopio
Canthia, Harjulantie 1 A, 1. krs.
70210 Kuopio, Finland
Tel: +358-017-162422
Fax: +358-017-162424
E-mail: Arturo.Garcia-Horsman at uku.fi


Vyaches Ibragimov wrote:

> Dear All.
> I use his-tag to purify the protein.
> But does anybody know if I can remove the his-tag after
> protein purification and how?
> Thanks..
> *******************************************************************************
> Slava Ibragimov, Ph.D. student
> Dept. of Life Sciences, Ben Gurion University
> Campus Beit Hias; Henrietta Szold St.; Bldg #4 Room 101;
> P.O.Box 653  Beer-Sheva 84105
> Israel
> Tel:972-7-6461360. Email:vyc at bgumail.bgu.ac.il
> *******************************************************************************
>
> ---




More information about the Methods mailing list