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Re: [ccp4bb] highly soluble proteins
 

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CCP4bb <-- 2007 <-- August 2007 <-- 07 August 2007
Previous message:
Subject: highly soluble proteins
From: Yvonne Leduc yal607 {- at -} MAIL {- dot -} USASK {- dot -} CA
Date: 2007-08-07 		Next message:
Subject: Postdoc position: Structural biology of intracellular signalling systems
From: Clemens Steegborn Clemens {- dot -} Steegborn {- at -} RUB {- dot -} DE
Date: 2007-08-07


Subject: Re: highly soluble proteins
From: Jack Tanner TannerJJ {- at -} MISSOURI {- dot -} EDU
Date: 2007-08-07
Try adjusting the pH close to the pI to reduce solubility.


> From: Yvonne Leduc
> Reply-To:
> Date: Tue, 7 Aug 2007 10:17:18 -0600
> To:
> Subject: [ccp4bb] highly soluble proteins
>
> Our lab is trying to crystallize a highly soluble (100+ mg/ml) protein
> with a molecular weight of 35 kd.
>
> The protein was screened against 1536 conditions at 20 mg/mL. Most drops
> were either clear or produced "bubbles" (often oily looking). The few
> that had precipitate contained high concentrations of K3PO4, cobalt, or
> zinc. We have tried repeating some of the bubble conditions at 100+
> mg/mL and are still getting clear drops or bubbles.
>
> Is there something about highly soluble proteins and/or secreted
> proteins and/or proteins with unusual portions of their sequence that
> needs to be considered in order to successfully crystallize it?
>
> I am considering trying "salting out" using dialysis, and also adding
> ligands/inhibitors. The protein is in 50 mM NaCl plus 50 mM buffer.
>
> I welcome thoughts and suggestions on crystallization ideas,
> publications, etc.
>
> Thank you
>
> Yvonne

CCP4bb navigation

CCP4bb <-- 2007 <-- August 2007 <-- 07 August 2007
Previous message:
Subject: highly soluble proteins
From: Yvonne Leduc yal607 {- at -} MAIL {- dot -} USASK {- dot -} CA
Date: 2007-08-07 		Next message:
Subject: Postdoc position: Structural biology of intracellular signalling systems
From: Clemens Steegborn Clemens {- dot -} Steegborn {- at -} RUB {- dot -} DE
Date: 2007-08-07


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