Dear Eike,
What is the interest of soaking experiments when you can grow HEWL crystals
in less time (15 min) than it would take to do a soaking experiment?
https://www.hamptonresearch.com/product_detail.aspx?cid=28&sid=173&pid=524
Soaking will work as long as you do it correctly. A 20min soak or longer
should work.
You should do it directly in the cryosolution with high sugar
concentrations as most diols might compete with the sugar.
Remember, as soon as you put crystals with bound GlcNac in a cryosolution,
you will start to back-soak the ligand.
Always add the GlcNac to the cryoprotectant even for co-crystals for the
best ligand density.
You may find the following reference useful to select a cryosolution that
does not dissolve your crystals:
Ciccone L., Vera L., Tepshi L., Rosalia L., Rossello A. & Stura E.A.
(2015) Multicomponent mixtures for cryoprotection and ligand
solubilization. Biotechnology Reports 7:120—127.
Enrico.
On Mon, 20 Feb 2017 20:59:23 +0100, Schulz, Eike-Christian
<eike.sch...@mpsd.mpg.de> wrote:
Dear all,
There are several structures in the PDB that show Lysozyme (mutants) in
complex with GlcNac (or similar compounds). However, all of those
structures seem to originate from co-crystallization experiments. I was
wondering whether anybody knew a successful case of complex formation by
soaking. But maybe the solvent channels are too small …
I would be very happy if anybody could point me to a reference but I
could also live with anecdotal evidence.
With best regards
Eike
--
Enrico A. Stura D.Phil. (Oxon) , Tel: 33 (0)1 69 08 4302 Office
Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab
LTMB, SIMOPRO, IBiTec-S, CE Saclay, 91191 Gif-sur-Yvette, FRANCE
e-mail: est...@cea.fr Fax: 33 (0)1 69 08 90 71
Proxima-2A, Soleil Synchrotron. Tel: 33 (0)1 69 35 8180 Beamline
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