Re: [ccp4bb] crystallizing fusion proteins

CP4BB@JISCMAIL.AC.UK" Subject: Re: [ccp4bb] crystallizing fusion proteins Hi Herman, We learned a few tricks of using MBP fusion for solving an interesting fold of a receptor extracellular domain from our recent publication (https://pubmed.ncbi.nlm.nih.gov/32541044/). (1) the length of a link

Re: [ccp4bb] crystallizing fusion proteins

Hi Herman, We learned a few tricks of using MBP fusion for solving an interesting fold of a receptor extracellular domain from our recent publication (https://pubmed.ncbi.nlm.nih.gov/32541044/ ). (1) the length of a linker between MBP and protein of

Re: [ccp4bb] [EXT] Re: [ccp4bb] crystallizing fusion proteins

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 <4aa44fc90f38-dmarc-requ...@jiscmail.ac.uk> wrote:  WARNING: This email originated from outside of MD Anderson.

Re: [ccp4bb] crystallizing fusion proteins

For us, a C-terminal macro domain tag worked well to crystallize a rather flexible target protein. https://pubmed.ncbi.nlm.nih.gov/27774698/ best regards, Michael On 15.03.21 16:51, Pascal Egea wrote: Hi Herman, I will add a few points to all the excellent advice already provided to you.

Re: [ccp4bb] crystallizing fusion proteins

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

Re: [ccp4bb] crystallizing fusion proteins

343 Royal Parade Parkville, VIC, 3052 +613 9662 7304 +614 57 539 419 tom.p...@csiro.au From: CCP4 bulletin board on behalf of Schreuder, Herman /DE Sent: Monday, March 15, 2021 8:07 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] crystallizing fusion proteins

Re: [ccp4bb] crystallizing fusion proteins

If you have a biophysical way to confirm functionality and/or folding it would go a long way towards helping you make this choice. Does your domain bind a ligand of any kind? Is it significantly stable in a simple thermal melt experiment (of course you have to account for the melting of the

[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