Re: [ccp4bb] crystallizing fusion proteins

Hi Herman I agree with all discussed already. First, is your protein of interest actually folded or just solubilized by the large fusion partner? If it is folded, worth considering crystallization of the fusion protein, as a “desperate measure” approach. The key issue will be making the

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

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 There are published reports of proteins crystallising with fusion partners, but I suspect there are a whole lot of unpublished results of

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

[ccp4bb] Research Fellow position at A*STAR in Singapore

*Research Fellow Position in Structural Biology* *Date available: immediately* *Date posted: 15 March 2021* We are seeking an enthusiastic and self-motivated postdoctoral research fellow to work in laboratory of structural biology for signalling and diseases in Institute of Molecular and