Hello Marco,
I know you said you would prefer keeping the full-length protein. But have you
looked at an AlphaFold2 prediction? Ideally of the monomer and homo- and
hetero-dimers you mentioned. It’s often a good guide for construct design. If
you see a lot of “spaghetti” with low pLDDT, that mi
Adding to the excellent advice offered by others:
1. Try to purify a unique oligomeric state: mixtures of homodimers,
heterodimers etc are less likely to crystallize. Collect a narrow peak off a
size exclusion column.
2. Try adding mono- or dinucleotides during crystallization, with or without
d
I'll second limited proteolysis with chymotrypsin.
Best wishes, Jon Cooper.
jon.b.coo...@protonmail.com
Sent from Proton Mail Android
Original Message
On 10/01/2025 07:02, Jeroen Mesters wrote:
> Hi,
>
> three things come to mind besides what has been mentioned already
>
> - m
stabilising your protein
(if this is an issue).
Cheers,
Sara
From: CCP4 bulletin board on behalf of Marco Bravo
<d0eb7bee83ae-dmarc-requ...@jiscmail.ac.uk>
Sent: 10 January 2025 01:00
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Help crystalizing protein
C
Hi,
three things come to mind besides what has been mentioned already
- many proteins that have an affinity for nucleic acids actually love sulfates
or phosphates… And yes, phosphates may lead to salt crystals at higher pH
values but this should not stop you a priori from screening with the sam
Hi Marco,
I wonder what is the highest concentration of the protein you've tried.
Once I worked with a protein which would crystallize only if it was
concentrated into a jell...
Good luck!
Nukri
On Thu, Jan 9, 2025 at 7:01 PM Marco Bravo <
d0eb7bee83ae-dmarc-requ...@jiscmail.ac.uk> wrote:
> H
Hello and you are welcome.
- You can add things after you purify, or buffer exchange on a concentrator
- sometimes one residue change makes a difference, removing 20 is almost
certainly going to change things, maybe for the better
- fusion of guest domains was mentioned with the intent of crysta
Hello Marco
This is a classic case of protein that just won't behave :)
Nil desperandum!
Without rewriting several large ish chapters on protein crystallization,
let's review your options from a bird's eye view:
0. Modify crystallization setup
This is something that you already have explored co
hey! I would like some more information... which buffers have you used and
at what pH?
Em qui., 9 de jan. de 2025, 22:01, Marco Bravo <
d0eb7bee83ae-dmarc-requ...@jiscmail.ac.uk> escreveu:
> Hello all,
> I am seeking some advice or ideas to crystalize a nuclease protein. It
> exists within a
