On Wed, Feb 1, 2012 at 11:17 AM, Dianfan Li <[log in to unmask]> 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 (domain closure) when binding ATP.
> 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?
I'd recommend trying ATP-gammaS - it could also be a substrate, but
it's worth a look. (Is there any reason to believe that AMP-PNP is a
substrate?) I've noticed that the various analogues have been known
to result in different conformations in the crystal structure, so it
may be a good idea to try more than one anyway.
-Nat
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