You may want to have a look at 3ur7 and 3ur8. I have also another example of a glycohydrolytic enzyme with an N-terminal His-tag sitting in the active site of a symmetry-related molecule (not deposited yet).
Karolina On 27/6/2012, "Brad Bennett" <[email protected]> wrote: >I think it was an N-terminal RGS-type His tag in 3O8Y (human lipoxygenase) >that mediated crystal contacts with a symmetry related molecule. As I >recall, this tag composed a B-strand that formed a nice interface with a >"native" B-strand of the symmetry related molecule. Pretty cool... > >-Brad > >On Wed, Jun 27, 2012 at 11:00 AM, Phoebe Rice <[email protected]> wrote: > >> With Flp recombinase - DNA complexes, a C-terminal His tag triggered a >> different (but sadly not better) crystal form, and the His side chains >> packed against the bases at the end of a neighboring DNA duplex. >> >> ===================================== >> Phoebe A. Rice >> Dept. of Biochemistry & Molecular Biology >> The University of Chicago >> phone 773 834 1723 >> >> http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123 >> http://www.rsc.org/shop/books/2008/9780854042722.asp >> >> >> ---- Original message ---- >> >Date: Wed, 27 Jun 2012 10:14:58 -0400 >> >From: CCP4 bulletin board <[email protected]> (on behalf of "R. M. >> Garavito" <[email protected]>) >> >Subject: Re: [ccp4bb] The effect of His-tag location on crystallization >> >To: [email protected] >> > >> > Most of the comments you will get will be anecdotal >> > in that people will report the successful results >> > and do not take the time or effort to characterize >> > the less successful results. This often occurs >> > because the tagged portion of the protein is most >> > often disordered, even in the best crystals. Thus, >> > other than saying "tagging on this end works, but >> > tagging on that end doesn't," there is little more >> > you can say. Each case will be different, and it is >> > almost impossible to arrive at any generalized >> > conclusion. >> > We prefer C-terminal tagged proteins for a number of >> > reasons, but if an N-terminally tagged protein >> > crystallizes well, so be it. Of the dozens of N- >> > and C-tagged protein structures we have solved in my >> > lab and with collaborators, I have only seen one >> > case of an ordered His-tag: the His residues had >> > coordinated Cd ions, which proved essential for >> > getting good crystals. However, beyond that there >> > was not much more to say. >> > For your protein and the resulting crystals, an >> > N-terminally tagged protein crystallized well. >> > Whether you can draw any more conclusions from >> > these results depends on characterizing crystals of >> > both N- and C-tagged proteins. Just assuming that >> > the C-tagged protein is trying to crystallize in the >> > same or related crystal form as the N-tagged protein >> > is an unwarranted assumption without experimental >> > evidence to back it up. That is why most groups >> > just run with the winner. >> > Cheers, >> > Michael >> > **************************************************************** >> > R. Michael Garavito, Ph.D. >> > Professor of Biochemistry & Molecular Biology >> > 603 Wilson Rd., Rm. 513 >> > Michigan State University >> > East Lansing, MI 48824-1319 >> > Office: (517) 355-9724 Lab: (517) 353-9125 >> > FAX: (517) 353-9334 >> > Email: [email protected] >> > **************************************************************** >> > On Jun 26, 2012, at 9:06 PM, weliu wrote: >> > >> > Dear all, >> > >> > We crystallized a protein and found that crystal >> > quality greatly depended on the location of >> > His-tag. When a His-tag was added at the >> > C-terminus, only crystalline precipitate or >> > spherical quasi crystals were grown. However, when >> > the His-tag was moved to the N-terminus, single >> > crystals were grown under a number of conditions, >> > and the best one diffracted to 1.7 angstrom after >> > optimization. I was wondering if there were >> > published reports describing similar cases. >> > >> > Thank you in advance >> > >> > Wei Liu >>
