Message-
From: CCP4 bulletin board on behalf of Guenter Fritz
Sent: Fri 9/26/2008 3:21 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] losing zinc during crystallization
Hi Sue,
I fully agree with Thierry, you might have to cyrstallize the protein
under exclusion of dioxygen
: Re: [ccp4bb] losing zinc during crystallization
Here is my two cents...
How strong zinc is captured by the protein is very protein dependent: I
always thought that a great case for this variability was the protection
of RING ubiquitin ligases against NEM. Cysteine-catalyzing HECT
ubiquitin
Hi Sue,
I fully agree with Thierry, you might have to cyrstallize the protein
under exclusion of dioxygen. There are many metallo proteins which have
to be crystallized that way. But a first attempt might be the TCEP as
already suggested by many others. If you need advice regarding
Hello Everyone
I've been trying to crystallize a zinc-containing enzyme for what
seems to me to be an eternity. The protein contains stoichiometric
zinc (1 zinc/ protein monomer) when isolated and the zinc is required
for activity. Each crystal we've obtained has lost the zinc and
Based on my own experience with zinc-metalloenzymes with thiolate
ligands, it's usually more a problem to get the zinc OUT than get it IN.
Zinc is pretty thiophilic, so removing it once ligated in a multiple Cys
environment is often difficult. Have you tried TCEP as a reducing agent
for
We had a zinc-finger containing protein that we were soaking with different
heavy atom compounds. It turns out KAu(CN)2 provided the best diffraction
of several soaks. We found out it was because the gold had replaced the
zinc ion and was coordinated by the Cys and His's. The lab nickednamed
I don't know if anyone had experience of TCEP inducing zinc acetate to
precipitate. This paper mentioned this.
The Crystal Structure of the Olfactory Marker Protein at 2.3 Å Resolution
*
Paul C. Smith, Stuart Firestein and John F. Hunt
Journal of Molecular
or
purification? This would change some of the other solutions that you might
attempt.
Cheers,
Artem
-Original Message-
From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Sue
Roberts
Sent: Thursday, September 25, 2008 5:36 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] losing zinc during
Han-Chun Tsai
Sent: Thursday, September 25, 2008 6:18 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] losing zinc during crystallization
I don't know if anyone had experience of TCEP inducing zinc acetate to
precipitate. This paper mentioned this.
The Crystal Structure of the Olfactory
*Subject:* Re: [ccp4bb] losing zinc during crystallization
I don't know if anyone had experience of TCEP inducing zinc acetate to
precipitate. This paper mentioned this.
The Crystal Structure of the Olfactory Marker Protein at 2.3 Å Resolution
*Paul C. Smith, Stuart Firestein and John F. Hunt
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