[ccp4bb] highly soluble proteins

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CCP4bb <-- 2007 <-- August 2007 <-- 07 August 2007

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Subject: Re: needles vs crystals
From: Roger Rowlett rrowlett {- at -} MAIL {- dot -} COLGATE {- dot -} EDU
Date: 2007-08-07

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Subject: Re: highly soluble proteins
From: Jack Tanner TannerJJ {- at -} MISSOURI {- dot -} EDU
Date: 2007-08-07

Subject: highly soluble proteins
From: Yvonne Leduc yal607 {- at -} MAIL {- dot -} USASK {- dot -} CA
Date: 2007-08-07

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: Re: needles vs crystals
From: Roger Rowlett rrowlett {- at -} MAIL {- dot -} COLGATE {- dot -} EDU
Date: 2007-08-07

Next message:
Subject: Re: highly soluble proteins
From: Jack Tanner TannerJJ {- at -} MISSOURI {- dot -} EDU
Date: 2007-08-07