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Dear Simon, That most probably won't work. You will have to reconstitute the Fe/S cluster in your protein before crystallization, either by extracting it from another Fe/S protein (with the same cluster) or by assembling it in your protein with Fe/S cluster synthesizing enzymes (see e.g. articles by D.R. Dean & L. Zheng ). Where did the cluster go in the first place? Is the protein refolded from inclusion bodies? Did the cluster get lost through oxygen exposure? In the first case you should refold your protein in the presence of the cluster. Also make sure that your protein actually is folded (by CD, NMR, maybe. DLS) - I have seen "refolded" proteins that were random coils and soluble well over 20mg/ml. In the second case you have to work anaerobically and - if done right - shouldn't have any problems. Good luck, Jens Yue Li wrote: >*** For details on how to be removed from this list visit the *** >*** CCP4 home page http://www.ccp4.ac.uk *** > > >Dear All, > >I am trying to crystallize Fe-S cluster protein by using Apo protein, iron >and sulphur. Does anyone has this kind of experience? Thanks > >Simon > > > >
