[ccp4bb] Summary of : Dissolving Hydrophobic ligands for co-crystallization and assays

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: co-crystallization VS crystal soaking
From: " Rain Field
Date: 2010-05-17

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Subject: updated mtz file or original one in refmac5
From: Jay Pan ccp4pitt {- at -} GMAIL {- dot -} COM
Date: 2010-05-17

Subject: Summary of : Dissolving Hydrophobic ligands for co-crystallization and assays
From: Tuhin Bhowmick tuhin_b {- at -} PHYSICS {- dot -} IISC {- dot -} ERNET {- dot -} IN
Date: 2010-05-17

Dear friends,
here I'm summarizing all of your suggestions regarding dissolving a
hydrophobic ligand (in this case, containig benzoic acid moities) for
crystallization and biochemical assays:

1)Sodium salts of benzoic acid containing molecules can often be soluble
in water to high concentration.If material is not limiting, it is a
strong possibility to be explored.

2)In case of a ligand with a protonated group (like benzoic acid),
increasing the pH of the medium helps the group to donate proton, in turn
forming carboxylate anaion, which dissolves better.The need to elevate pH
will vary depending upon the ligand.

3)Using low mol. wt. PEGs, such as 200, 300, 350 etc. to dissolve the
ligand. PEG mono methyl ether 350, 550 can also be tried.

4) using mild detergents (DDM, OG etc.)

5)(For xtalization, not for activity....)
Keep prot concentration as low as 1 mg/ml, then add you ligand in low
concentration according to molar equivalence as req. After incubation
(for whatever time required), concentrate the solution, then use for
xtalization.Final conc. of DMSO shouldn't exceed 2 - 5%.

Thanks to all for your time,
I will try out and let know.
Regards,
Tuhin.

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CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: co-crystallization VS crystal soaking
From: " Rain Field
Date: 2010-05-17

Next message:
Subject: updated mtz file or original one in refmac5
From: Jay Pan ccp4pitt {- at -} GMAIL {- dot -} COM
Date: 2010-05-17