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Re: [ccp4bb] problem of crystallization
 

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CCP4bb <-- 2008 <-- May 2008 <-- 16 May 2008
Previous message:
Subject: Re: problem of crystallization
From: Jens-Christian Navarro Poulsen jnp {- at -} KEMI {- dot -} KU {- dot -} DK
Date: 2008-05-16 		Next message:
Subject: Phaser compile errors
From: Nathan I Nicely nathan {- dot -} nicely {- at -} DUKE {- dot -} EDU
Date: 2008-05-16


Subject: Re: problem of crystallization
From: Clemens Grimm Clemens {- dot -} Grimm {- at -} BIOZENTRUM {- dot -} UNI-WUERZBURG {- dot -} DE
Date: 2008-05-16
Hi Jennifer,

clear drops can still be brought to supersaturation by:

-transfer to wells with lower vapour pressure: exchange or add LiCl or other
concentrated salt solutions in the reservoir

-temperature gradient: place trays on ice/cold surface while hanging drops are
at higher temperature, this will draw more water out of the drops.

-transient supersaturation might be sufficient to trigger nucleation and
subsequent crystal growth: slightly open wells for a few minutes (or longer)
and close again.

-protein solubility also can strongly depend on temperature: move to 4°C etc.

... and why not using the 100mg/ml stock or concentrate even more?

Good luck,
Clemens


Quoting Jens-Christian Navarro Poulsen :

> Hi Jennifer
>
>
>
> I just want to draw your attention the following paper regarding methylation
> of Lysines, which reduces the solubility of their test proteins.
>
>
>
> Walter
>
> 2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum> TS, Meier C,
> Assenberg R, Au KF, Ren J, Verma A, Nettleship JE, Owens RJ, Stuart DI,
> Grimes JM.
>
>
>
> 2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum> Abstract
>
> Lysine methylation as a routine rescue strategy for protein crystallization.
>
> Structure. 2006 Nov;14(11):1617-22.
>
> PMID: 17098187 [PubMed - indexed for MEDLINE]
>
>
>
> Best regards,
>
>
>
> Jens-Christian Navarro Poulsen
>
> Dept of Chemistry, KU
>
>
>
>
>
> _____
>
> From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
> Jennifer Han-Chun Tsai
> Sent: 13. maj 2008 18:17
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: [ccp4bb] problem of crystallization
>
>
>
> Hi,
>
>
>
> This topic is not related to CCP4. I am having problem of crystallizing
> one protein. It's a pretty small protein with size around 15kDa. I have
> stock concentration around 100mg/mL. Crystallization plates I set up are
> with concentration of 10mg/mL, 30mg/mL and 50mg/mL. All the plates had been
> set up at least one week. Only around 5 wells per plate or less formed
> precipitation. The rest of wells are pretty clear still. Is there any
> suggestion for reducing protein solubility or increasing the chance of
> getting crystals?
>
>
>
> Thanks for your time,
>
> Jennifer
>
>
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