Dear Dianfan,
In some cases the ATP-lid of the kinase is blocking the active site in the crystal form. In those cases the only option is to try co-crystallisation.
Besides ATP and the homologs you mention you can also try ADP that as you will see in the PDB has been heavily used for kinases.
Best of luck
Sofia
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-----Original Message-----
From: Dianfan Li <[log in to unmask]>
Sender: CCP4 bulletin board <[log in to unmask]>
Date: Wed, 1 Feb 2012 19:17:03
To: <[log in to unmask]>
Reply-To: Dianfan Li <[log in to unmask]>
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 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
behavour in a same way as AMP-PCP. Is it a good idea of trying quick
soaks at high concentrations of AMP-PCP? Co-crystallization is another
option I have but AMP-PCP is a substrate of the kinase (with low
rate).
What are other ways of getting ATP analogues into a crystal?
Thanks for suggestions,
Dianfan
Dianfan Li, PhD
College of Biochemistry and Immunology
Trinity College Dublin
Dublin, Ireland.
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