We have had really outstanding success using trimethyllead and
samarium acetate as 'quick soak' derivatives. Both are soluble to 100 mM
in most crystallization solutions and have large anomalous signals at the
copper-kalpha wavelength used at most home sources. A quick 30 sec to 5
minute s
Or you could try crystallizing the protein in the presence of KI or
NaI, and collect some in-house SAD data. You could also try to boost
the concentration and number of ordered halide sites by quick soaking
the crystals with a higher concentration of the iodide salt.
In my limited experienc
Hi Amit,
Dr. Dauter has several excellent papers about solving crystal structures with
halide ions. Use this search string in PubMed (without the quotes) to retrieve them: "dauter [AUTH] AND halide". The anomalous signal of iodide can be detected using a home x-ray source but the anomalous sig
ECTED]
***
- Original Message -
From: amit sharma
To: CCP4BB@JISCMAIL.AC.UK
Sent: Thursday, September 25, 2008 6:03 AM
Subject: [ccp4bb] Quick-soak
Dear CCP4bbers,
I have a protein molecule(~9.0 kDa) that crystallized in the presence of
0.15M KBr and HEPES p
2008/9/25 amit sharma <[EMAIL PROTECTED]>
>
> Dear CCP4bbers,
> I have a protein molecule(~9.0 kDa) that crystallized in the presence of
> 0.15M KBr and HEPES pH 7.0. Since there is no homologuous structure present,
> I intend to perform heavy metal derivatization. I read some literature which
>
Hi Amit,
http://www.doe-mbi.ucla.edu/~sawaya/tutorials/Phasing/references.html
And the (IMHO) seminal heavy atom derivative reference:
Petsko, G.A., "Perparation of Isomorphous Heavy-Atom Derivatives"
Methods in Enzymology, Volume 114, , pages 147-157.
should give you all the info you need.
Th
Dear CCP4bbers,
I have a protein molecule(~9.0 kDa) that crystallized in the presence of
0.15M KBr and HEPES pH 7.0. Since there is no homologuous structure present,
I intend to perform heavy metal derivatization. I read some literature which
suggested that I could carry out quick soak with 0.5M So
Hi,
similarly, 30 sec soak with 0.5 M potassium iodide. But in this case,
the iodides bound specifically to a site where otherwise chloride
binds. These halide binding sites were totally unexpected ...
Best regards,
Dirk.
Am 26.09.2007 um 01:55 schrieb James Whisstock:
Hi - sorry - rath
Dear Derek and Tassos,
I normally start using the following compounds depending on pH of the
cryo-solution at 10 mM concentration and 10 min incubation time and
optimize towards lower concentrations and longer incubation times:
-ethyl mercuric phosphate (EMP)
-HgCl2
-SmCl3
-K2PtCl4
-K2PtCl6
Be
I also solved a structure on our home CuKa source by SIRAS with 20-60sec in
0.5-1M KI.
JPK
==Original message text===
On Tue, 25 Sep 2007 6:55:28 pm CDT James Whisstock wrote:
Hi - sorry - rather than iodine I meant to say we had had success with
Potassium Iodide (1M f
Hi - sorry - rather than iodine I meant to say we had had success with
Potassium Iodide (1M for 20 seconds)!
Cheers
James
[EMAIL PROTECTED] wrote:>
> Hi,
>
> I do not use their method as such - however, I love heavy atom soaks and
> do them any time I can, so I've got very similar experience
Hi,
I do not use their method as such - however, I love heavy atom soaks and
do them any time I can, so I've got very similar experiences in the past.
Heavy atoms can bind very quickly even from quite dilute solutions - the
quickest I've ever soaked (and got useful data) was sodium chloroplatinat
Hi Derek
We have had success with iodine - 20 seconds soak.
J
Derek Logan <[EMAIL PROTECTED]> wrote:>
> Hi Uwe,
>
> Just what I wanted to hear, and with a limited set of compounds too!
> Follow-up question: what are these 4-6 most successful compounds?
> Thanks also for the tips on monitoring
There's a nice databank out there in the world wild web:
http://www.sbg.bio.ic.ac.uk/had/
Juergen
Derek Logan wrote:
Hi Uwe,
Just what I wanted to hear, and with a limited set of compounds too!
Follow-up question: what are these 4-6 most successful compounds?
Thanks also for the tips on
er 25, 2007 12:13 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] "Quick soak" method
Hi,
I'd like to find out how successful the "quick soak" method for heavy atom
derivatisation proposed by Radaev and Sun:
Sun PD, Radaev S, Kattah M. Generating isomorphous heavy-ato
Hi Uwe,
Just what I wanted to hear, and with a limited set of compounds too!
Follow-up question: what are these 4-6 most successful compounds?
Thanks also for the tips on monitoring thw soaking
Derek
On Sep 25, 2007, at 13:23, Uwe Mueller wrote:
Dear Derek,
we are using the quick-soak me
You may also wish to take a look in Dauter and Nagem's quick cryo
soaking for introducing anomalous scattering.
1. Acta Crystallogr D Biol Crystallogr. 2001 Jul;57(Pt 7):996-1002.
2. Methods Enzymol. 2003;374:120-37.
3. Acta Crystallogr D Biol Crystallogr. 2005 Aug;61(Pt 8):1022-30
HTH
[ ]s
-
Dear Derek,
we are using the quick-soak method frequently at the BESSY-MX beamlines.
So far we were able to obtain phase information in terms of MAD/SAD-
phases in all cases which we tried. The starting set-up is the screen
over the 4-6 most successful heavy atom compounds at 5-10mM
concentrat
Hi,
I'd like to find out how successful the "quick soak" method for heavy
atom derivatisation proposed by Radaev and Sun:
Sun PD, Radaev S, Kattah M. Generating isomorphous heavy-atom
derivatives by a quick-soak method. Part I: test cases. Acta Cryst.
2002. D58:1092-1098.
has been in co
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