Hi Armando, Have you looked at predicted models e.g. Alphafold for the full-length protein?
+ Are there experimentally determined structures for full length structural homologues or orthologues from other species to validate computational models? You may be able to align your domains against the full length that way and add-in the Alphafold model of the linker to generate a complete structure If there is significant contact surface between the two domains, then you may need to do some molecular dynamics to resolve clashes / create optimal inter-domain contacts – again the predicted model and / or homologous experimentally determined structures may help with seeing which bits ae likely to have conformational change in the complete structure vs the individual domain structures. Cheers, Charlie. From: CCP4 bulletin board <[email protected]> On Behalf Of Armando Albert Sent: 26 July 2023 09:14 To: [email protected] Subject: [ccp4bb] On the production of a two domain protein WARNING: This message was sent by an external party. Report suspicious messages via the “Report Phishing” button in Outlook. Dear all, We are trying to characterize a protein consisting of two domains. We have successfully produced, purified and crystallized both domains independently. However, we are unable to overexposes a soluble form of the full-length protein. Can anyone provide a strategy to merge back the independent domains into a single protein chain or to modify the construct to succeed in the overproduction of the full-length protein? Armando ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1<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<http://www.jiscmail.ac.uk/CCP4BB>, a mailing list hosted by www.jiscmail.ac.uk<http://www.jiscmail.ac.uk>, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/<https://www.jiscmail.ac.uk/policyandsecurity> ######################################################################## 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/
