Hi all, I am working on a newly identified E3 ubiquitin ligase (RING type). I am interested to co-crystallize it with E2 enzyme. Among the papers reported so far, which are just a few, people have used a mixture of E2s including UbcH1, UbcH5 and UbcH6 for the ubiquitination assays. However, there is no clear evidence on E2 enzyme functions with this E3 ubiquitin ligase in vivo.
Can someone suggest how can I choose one particular E2 (among UbcH1/UbcH5/UbcH6/UncH7...) for co-crystallization studies. It should be noted that all these E2 enzymes have a similar fold, so will it make sense to just randomly choose any one of these E2 enzymes and get started. Also, if someone with experience in this area suggest weather a Ub-E2-E3 complex is stable enough and suitable for ternary complexes. Thanks in advance. -Geet
