As others suggested, I would check the globularity of the fusion protein with biophysical technique such as SAXS, if possible.
Aleem On Mar 15, 2021, at 10:52 AM, Pascal Egea <00004aa44fc90f38-dmarc-requ...@jiscmail.ac.uk> wrote: WARNING: This email originated from outside of MD Anderson. Please validate the sender's email address before clicking on links or attachments as they may not be safe. Hi Herman, I will add a few points to all the excellent advice already provided to you. If you decide to try this option of crystallizing a fusion protein of your target, I would consider using N-terminal but also C-terminal fusions. We have had success using MBP in N-ter and the superfolder GFP in C-terminal for a few 'vexing' proteins from Plasmodium falciparum. It helped us either to obtain crystals/structures (as you seek) but also provided a way to circumvent a tendency for twinning from our target in some specific cases. GFP has the added benefit that your fusion crystals should be bright yellow so that speeds up the screening process a bit. I hope this helps. Good luck Best regards, Pascal Egea, PhD UCLA School of Medicine ________________________________ 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://urldefense.com/v3/__https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1__;!!PfbeBCCAmug!zZxYLQsNYEE7cGG5xqck4mGlLhxy_RFp_9C-98qrEOdMHkcrQeVJ0b4Mrb0GrUXvIw$> <https://urldefense.com/v3/__http://mednet.ucla.edu__;!!PfbeBCCAmug!zZxYLQsNYEE7cGG5xqck4mGlLhxy_RFp_9C-98qrEOdMHkcrQeVJ0b4Mrb00MzwdbQ$> ________________________________ 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://urldefense.com/v3/__https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1__;!!PfbeBCCAmug!zZxYLQsNYEE7cGG5xqck4mGlLhxy_RFp_9C-98qrEOdMHkcrQeVJ0b4Mrb0GrUXvIw$> The information contained in this e-mail message may be privileged, confidential, and/or protected from disclosure. This e-mail message may contain protected health information (PHI); dissemination of PHI should comply with applicable federal and state laws. If you are not the intended recipient, or an authorized representative of the intended recipient, any further review, disclosure, use, dissemination, distribution, or copying of this message or any attachment (or the information contained therein) is strictly prohibited. If you think that you have received this e-mail message in error, please notify the sender by return e-mail and delete all references to it and its contents from your systems. ######################################################################## 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/
