Hello Marco,

I know you said you would prefer keeping the full-length protein. But have you 
looked at an AlphaFold2 prediction? Ideally of the monomer and homo- and 
hetero-dimers you mentioned. It’s often a good guide for construct design. If 
you see a lot of “spaghetti” with low pLDDT, that might explain the low rate of 
crystallization hits you observed, and you might have better luck with all this 
truncated.

If you are trying to crystallize the complex with DNA, try the set of 
conditions described in this paper: https://doi.org/10.1107/S1744309112025316
(Although you would probably need a catalytically dead mutant of the nuclease 
for this to be tractable.)

Good luck with your project!

Guillaume

On 13 Jan 2025, at 11:40, Opher Gileadi <ogile...@gmail.com> wrote:

Adding to the excellent advice offered by others:
1. Try to purify a unique oligomeric state: mixtures of homodimers, 
heterodimers etc are less likely to crystallize. Collect a narrow peak off a 
size exclusion column.
2. Try adding mono- or dinucleotides during crystallization, with or without 
different divalent cations.

Good luck

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