On Wed, Feb 1, 2012 at 11:17 AM, Dianfan Li <l...@tcd.ie> 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
