Dear all,
Thanks a lot for good suggestions on this topic. To answer some of the
questions: The apo strucuture has not been solved; the crystals dissolved upon
soaking (my term 'crack' was not accurate); and I have not tried
co-crystallization.
A brief summerization here:
1) Soak/Co-crystalli
Consider cross-linking crystals with glutaraldehyde. The caveat here is
that you may end up with the protein conformation that is forced by
lattice, but if the issue is just the fragility, you should be fine. I
assume that crystals simply crack but do not dissolve?
Certainly, as others have said
Maybe someone has suggested this already... If so, I am re-enforcing it.
If the cracking is coming from actual molecular movement induced by binding
(and not other reason like differing ionic strength in your soaking conditions)
you could try setting up some co-crystallization and (hopefully)
gr
.
Best of luck
Sofia
Sent via BlackBerry® from Vodafone
-Original Message-
From: Dianfan Li
Sender: CCP4 bulletin board
Date: Wed, 1 Feb 2012 19:17:03
To:
Reply-To: Dianfan Li
Subject: [ccp4bb] Soaking Kinase Crystals with ATP analogues
Dear all,
Sorry about
Hi ,
To add to the previous comments,
crystallization of GTP or ATP (or their analogues) with their kinase/ A- or
G-tpases can depend on a lot of factors that were mentioned (such as
packing).
A simple common problem is that ATP solutions should be carefully buffered
prior to their use for soakin
972-8-646-1710
From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Dianfan Li
[l...@tcd.ie]
Sent: Wednesday, February 01, 2012 9:17 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Soaking Kinase Crystals with ATP analogues
Dear all,
Sorry about a non-crystallographic quest
I assume that cocrystallization has failed? What you are experiencing is
likely the effect of conformational transition caused by ligand. You can
try very slow adition (even microdialysis) or if your ligand is fairly
insoluble then you can just add a tiny solid particle of inhibitor to your
drop an
On Feb 1, 2012, at 12:17 PM, Dianfan Li wrote:
> I am working on a kinase and would like to get an ATP analogue into
> the crystals. When soaked with AMP-PCP, the kinase crystals crack in
> about 15 min at 4 C.
15 minutes is a long time. Scoop crystals during that time period.
Do the cracked
etin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Dianfan Li
Sent: Wednesday, February 01, 2012 2:17 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Soaking Kinase Crystals with ATP analogues
Dear all,
Sorry about a non-crystallographic question here.
I am working on a kinase and would like to get an ATP a
On Wed, Feb 1, 2012 at 11:17 AM, Dianfan Li wrote:
> I am working on a kinase and would like to get an ATP analogue into
> the crystals. When soaked with AMP-PCP, the kinase crystals crack in
> about 15 min at 4 C.
This isn't too surprising; most kinases undergo global conformational
changes (dom
Dear all,
Sorry about a non-crystallographic question here.
I am working on a kinase and would like to get an ATP analogue into
the crystals. When soaked with AMP-PCP, the kinase crystals crack in
about 15 min at 4 C.
I could try other analogues like AMP-PNP etc, but those would probably
behavou
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