Hello all, Does anyone know of an example where SHARP was used to phase multiple sulphur-SAD datasets and combine all of the data into one map?
I have multiple crystals of the same protein, but each crystal form has a different LtoM mutation (the sulfur content of the native construct is too low for phasing without the mutations). The crystals are sensitive to oxidaton, so I cannot make one construct with all of the mutations, and selenomethionine incorporation is out due to the oxidaton issues. Each crystal form from all mutants is isomorphous. I would like to collect highly redundant datasets of each form, and then use SHARP to find all the Sulphurs, and combine the initial phases from all datasets with the native data and build the model. The protein has two domains, and I know the structure of one of them (~40% of the total), so I should be able to use the intial MR phases to help with finding the sulphurs.... Is this feasible? Are there any examples like this out there? Could something like ths also be done in MLPHARE or CRANK, etc? I want to make sure this is sound before I head off to a synchrotron with a million crystals! Thanks everyone!
