Hi all,
I've got crystals of a protein of ca 200 residues, with 2 free
cysteines, 5 histidines, 2 methionines.
We have nice diffraction for the native crystals, that grow in 150 mM
KSCN, 17% PEG 3350, bis tris propane pH 8.8.
We are crystallising the SeMet derivative, but I'm not completely su
Hi Sebastiano,
Have a look at HATODAS:
http://hatodas.harima.riken.go.jp/
Good luck,
Miguel
Le 15 juil. 09 à 14:33, Sebastiano Pasqualato a écrit :
Hi all,
I've got crystals of a protein of ca 200 residues, with 2 free
cysteines, 5 histidines, 2 methionines.
We have nice diffraction for
Hi Sebastiano,
Free Cys' are crying out for Mercury derivatives. Try 2-3 with varying
sizes of additional groups - HgCl2, K2HgI4, PCMB, from the "Magic
seven" would be a good place to start.
Watch out for problems with your low pH, though. Some salts will form
insoluble hydroxide salts at pH 8.8
luck
Savvas
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of David
Briggs
Sent: Wednesday, July 15, 2009 3:27 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] heavy atom derivative choice
Hi Sebastiano,
Free Cys' are crying out for Me
Whps!
Watch out for problems with your *HIGH* pH, though. Some salts will form
insoluble hydroxide salts at pH 8.8. (HgCl2 & K2HgI4 should be okay).
2009/7/15 Sebastiano Pasqualato :
> Hi all,
> I've got crystals of a protein of ca 200 residues, with 2 free cysteines, 5
> histidines, 2 methi
Hi,
Mercury will likely form a precipitate with you SCN- anions...
solubility in water is 0.6 g/L (about 3 mM) at 25°C...
Since you are not in water, at less than 25°C, and with you high
concentration of SCN- ions, don't expect to have high concentrations of
*soluble* Hg++ in you medium. Prolo
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Sebastiano Pasqualato
Sent: Wednesday, 15 July, 2009 14:34
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] heavy atom derivative choice
Hi all,
I've got crystals of a protein of ca 200 residues, with 2
Dear Sebastiano,
why not have a look at
http://sis.niaid.nih.gov/cgi-bin/heavyatom_reactivity.cgi.
A reprint of the original paper can be found at
http://sis.niaid.nih.gov/pub_pdf/Agniswamy-et-al-2008.pdf
We did not specifically look at Bis-Tris Propane but did look at Tris pH 8.5
and H
...@gsk.com
"Miguel Ortiz Lombardia"
Sent by: "CCP4 bulletin board"
15-Jul-2009 09:21
Please respond to "Miguel Ortiz Lombardia"
To
CCP4BB@JISCMAIL.AC.UK
cc
Subject
Re: [ccp4bb] heavy atom derivative choice
Hi Sebastiano,
Have a look at HATODAS:
htt
-isomorphism issues.
Best of luck
Savvas
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf
Of David
Briggs
Sent: Wednesday, July 15, 2009 3:27 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] heavy atom derivative choice
Hi Sebastiano,
Free Cys'
I would also include a NaI and NaBr quick soak in 250 mM, SCN- is
considered a pseudohalide
cheers Preben
Sebastiano Pasqualato wrote:
Hi all,
I've got crystals of a protein of ca 200 residues, with 2 free
cysteines, 5 histidines, 2 methionines.
We have nice diffraction for the native crystals
Dear All,
We have a very similar problem, I have got a protein of 200 residues
predicted to have two free cys (but 14 internal cys!), 1 free histidine, 2
methionines (all predicted to be free).
Native crystals diffracted to 2.2 A was obtained in 200 mM KSCN, 20% PEG
3350, 13% glycerol, bis t
X Xiong, Cellular & Molecular Medicine wrote:
My Question is:
Does mercury tends to get into the protein core to denature protein or not?
This is more likely to happen for a small "bare" Hg like Hg2+ in HgCl2
or Hg(OAc)2 than it is for a large organomercury compound like PCMB,
PCMBS etc so i
Wednesday, July 15, 2009 11:32 AM
Subject: Re: [ccp4bb] heavy atom derivative choice
X Xiong, Cellular & Molecular Medicine wrote:
My Question is:
Does mercury tends to get into the protein core to denature protein or
not?
This is more likely to happen for a small "bare" Hg l
Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: j-kell...@northwestern.edu
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- Original Message -
From: "Phil Jeffrey"
To:
Sent: Wednesday, July 15, 2009 11:32 AM
Subject: Re: [ccp4bb] heav
Hi,
1. KSCN will suck up a lot of the Hg and other metals -- can you switch to
something else?
2. Hg typically does not enter the protein core.
3. It's generally not necessary to use a special tank, as long as you
treat the resulting solutions as heavy-atom waste; but of course each
lab's safety p
>For gel-shift assay, do people normally use a special gel tank for heavy
>metal work?
We used to use a Pharmacia Phast system, as this is bufferless and made it
very easy to contain heavy metal contamination. Ours caught fire and died a
couple of years ago, and I have yet to find a good cheap r
David Briggs schrieb:
Bart Hazes used to have an excellent set of notes online about heavy
atom derivatisation - I can't seem to find the URL right now...
I believe the CCP4 wiki has the information that you're looking for (at
http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Soak
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