Hello Herman, If you have plenty of the fusion protein, there is probably no reason not to set it up in crystallisation trials and see if you get something. There are published reports of proteins crystallising with fusion partners, but I suspect there are a whole lot of unpublished results of this not working as well. As Artem has already stated, it could be that your protein of interest just doesn't like the buffer conditions when it finds itself alone in solution after cleavage, so that is an area to explore. Best of luck, tom
Tom Peat, PhD Proteins Group Biomedical Program, CSIRO 343 Royal Parade Parkville, VIC, 3052 +613 9662 7304 +614 57 539 419 tom.p...@csiro.au ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Schreuder, Herman /DE <herman.schreu...@sanofi.com> Sent: Monday, March 15, 2021 8:07 PM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: [ccp4bb] crystallizing fusion proteins Dear Bulletin Board, Sorry for the slightly off-topic question, but we are struggling with a receptor domain that expresses well as a fusion protein, but gets lost the moment it is cleaved from the fusion partner. It could be that the receptor domain is not or misfolded, but it could also be a solubility problem. I have seen some crystal structures of fusion proteins with MBP and for membrane proteins, T4-lysozyme fusions are often crystallized. What is your experience? Would it be worth trying to express and crystallize a fusion protein, or would it be better to look for other constructs, e.g. to include more receptor domains? Thank you very much for your advice! Herman ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ######################################################################## 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/
