Re: [ccp4bb] Zn fingers and Ni columns

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: Re: Gordon conference on membrane protein structure/function
From: Susan Buchanan skbuchan {- at -} HELIX {- dot -} NIH {- dot -} GOV
Date: 2008-01-10

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From: Dale Tronrud det102 {- at -} UOXRAY {- dot -} UOREGON {- dot -} EDU
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Subject: Re: Zn fingers and Ni columns
From: Kendall Nettles knettles {- at -} SCRIPPS {- dot -} EDU
Date: 2008-01-10

Phoebe,
We were able to purify the estrogen receptor DNA binding domain with a
6his-tag. After cutting off the tag and re-applying to the Ni-NTA matrix,
the protein did not stick to the beads. We had some low resolution crystals
that contained a second protein and DNA, which suggests to me that the zinc
was bound in the protein, but we did not analyze it further.

Kendall
--
Kendall W. Nettles, PhD
Assistant Professor
Department of Cancer Biology
The Scripps Research Institute
5353 Parkside Dr.
Jupiter Fl 33458

office 561-799-8851
fax 561-799-8805
cell 561-306-7566

On 1/3/08 7:21 PM, "price@UCHICAGO.EDU" wrote:

> This has probably been discussed before, so apologies in advance.
> We're eyeing a protein that has a probable C4 Zn finger in the
> middle. The collaborators who are nicely going to PCR it up want to
> know if we'd like it with or without a His tag.
> Is it a bad idea to co-mingle Zn-binders and Ni columns? Or is it
> likely to bind the column quite nicely without the tag?
> thanks,
> Phoebe
>
>
> ------------------------------------------------------------------------------
> ---------------------------------------------
> Phoebe A. Rice
> Assoc. Prof., Dept. of Biochemistry & Molecular Biology
> The University of Chicago
> phone 773 834 1723
> fax 773 702 0439
> http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabe
> tically.php?faculty_id=123
> http://www.nasa.gov/mission_pages/cassini/multimedia/pia06064.html

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Gordon conference on membrane protein structure/function
From: Susan Buchanan skbuchan {- at -} HELIX {- dot -} NIH {- dot -} GOV
Date: 2008-01-10

Next message:
Subject: Re: unbiased electron density map
From: Dale Tronrud det102 {- at -} UOXRAY {- dot -} UOREGON {- dot -} EDU
Date: 2008-01-10