I am working on two proteins of the UPS, that have been shown to interact with each other in vivo. The ultimate goal is to study the complex using X-ray crystallography. I would also like to study if this interaction is direct or mediated by other protein partners.

So far I have not been successful in establishing the interaction in vitro. If I purify the proteins separately and put them together for crystallization, will this help in anyway?

I have also tried performing Co-immunoprecipitation experiments in vivo and check for potential interaction partners using MS analysis, but this has not panned out either.

I would be grateful if anyone has inputs or suggestions as to how i can take this ahead. Thanks in advance!

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