Re: [ccp4bb] question about zinc binding protein

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

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From: Nathaniel Echols nathaniel {- dot -} echols {- at -} GMAIL {- dot -} COM
Date: 2010-03-31

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Subject: B factor Coloring in Pymol
From: "Clayton, Gina Martyn" gina_clayton {- at -} MERCK {- dot -} COM
Date: 2010-03-31

Subject: Re: question about zinc binding protein
From: Enrico Stura estura {- at -} CEA {- dot -} FR
Date: 2010-03-31

Not strange at all:

Reading from:

http://www.chem.gla.ac.uk/research/groups/protein/mirror/stura/cryst/add.html

"Zinc acetate or sulfate 0.2-5mM It inevitably reduces protein solubility.
It can act as an inhibitor. Essential for activity of various enzymes"

Zinc reduces protein solubility and often aggregates proteins. In a case
of a zinc binding protein you may need to add 100-200 mM imidazole to
control even 0.2mM concentrations of this ion.
Without good control, you will end up by depleating the zinc within the
precipitate
and end up without zinc in your binding site.

This is what you got.

Enrico.

On Wed, 31 Mar 2010 18:11:46 +0200, zq deng wrote:

> hello everyone, recently i purify a protein conteining zinc binding
> domain,and i want to determine its structure.i get the crystal,but poor
> diffraction.so i try to adding zinc into the protein to optimize the
> crystal,but the protein precipitate immidiately even the znic is 1
> mM.BTW,we use the protein to do zinc scan,we don't find the zinc. does
> anyone have some advice?hello everyone, recently i purify a protein
> conteining zinc binding domain,and i want to determine its structure.i
> get
> the crystal,but poor diffraction.so i try to adding zinc into the
> protein to
> optimize the crystal,but the protein precipitate immidiately even the
> znic
> is 1 mM.BTW,we use the protein to do zinc scan,we don't find the zinc.
> does
> anyone have some advice?

--
Enrico A. Stura D.Phil. (Oxon) , Tel: 33 (0)1 69 08 4302 Office
Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab
LTMB, SIMOPRO, IBiTec-S, CEA Saclay, 91191 Gif-sur-Yvette
Cedex FRANCE http://www-dsv.cea.fr/en/ibitecs/82
http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html
e-mail: estura@cea.fr Fax: 33 (0)1 69 08 90 71

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

Next message:
Subject: B factor Coloring in Pymol
From: "Clayton, Gina Martyn" gina_clayton {- at -} MERCK {- dot -} COM
Date: 2010-03-31