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 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/ VARNING: Klicka inte på länkar och öppna inte bilagor om du inte känner igen avsändaren och vet att innehållet är säkert. CAUTION: Do not click on links or open attachments unless you recognise the sender and know the content is safe. När du har kontakt med oss på Uppsala universitet med e-post så innebär det att vi behandlar dina personuppgifter. För att läsa mer om hur vi gör det kan du läsa här: http://www.uu.se/om-uu/dataskydd-personuppgifter/ E-mailing Uppsala University means that we will process your personal data. For more information on how this is performed, please read here: http://www.uu.se/en/about-uu/data-protection-policy ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
