Re: [ccp4bb] Crystallization of lysine and arginine rich proteins

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

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- X-ray detectors

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

Previous message:
Subject: Re: Crystallization of lysine and arginine rich proteins
From: "gohlke, ulrich" ulrich {- dot -} gohlke {- at -} MDC-BERLIN {- dot -} DE
Date: 2009-11-02

Next message:
Subject: Re: Crystallization of lysine and arginine rich proteins
From: Enrico Stura estura {- at -} CEA {- dot -} FR
Date: 2009-11-02

Subject: Re: Crystallization of lysine and arginine rich proteins
From: Andy Torelli att29 {- at -} CORNELL {- dot -} EDU
Date: 2009-11-02

Umar,

Check out: Czepas et al. The impact of Lys-->Arg surface mutations on
the crystallization of of the globular domain of RhoGDI, Acta D (2004)
60 275-280. They point out that sulfate ions can help mediate contacts
between arginine residues from neighboring molecules in the crystal.

You may have already considered the surface entropy server
(http://nihserver.mbi.ucla.edu/SER/) to help identify any specific
stretches of amino acids that could be mutated to reduce entropy and
possibly promote crystallization. Maybe there are a couple regions of
your sequence that are flagged as particularly unfavorable in terms of
their predicted entropy contribution. There is evidence that lysine is
more of a problem in this respect as compared to arginine (Derewenda et
al. Acta D (2006) 62 116-124).

Good luck,
-Andy

===========================================
Andrew T. Torelli Ph.D.
Postdoctoral Associate
Department of Chemistry & Chemical Biology
Baker Laboratory, Cornell University
Ithaca, NY 14853
===========================================

On 11/2/2009 8:39 AM, Jan Rash wrote:
> Dear All,
>
> I have a question regarding the crystallization of lysine and arginine
> rich protein around 13%. So far our attempts to crystallize this protein
> have not been successful although the secondary structure predictions,
> CD spectroscopy measurements clearly show that this protein is folded. I
> presume that these lysine and arginine are the sources of the local
> flexibility in the protein even though the protein is globular overall.
> Moreover, my attempts to crystallize the limited proteolysis fragments
> also did not achieve crystals. I have also tried the crystallization
> with its binding partners and could not succeed. I think any compound
> that binds to the lysine/arginine side chains might affect the
> crystallization process thereby reducing the internal flexibility of
> protein. Can anybody suggest some effective strategy for the
> crystallization?
>
>
>
> Thanks
>
> Umar
>
>
>

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

Previous message:
Subject: Re: Crystallization of lysine and arginine rich proteins
From: "gohlke, ulrich" ulrich {- dot -} gohlke {- at -} MDC-BERLIN {- dot -} DE
Date: 2009-11-02

Next message:
Subject: Re: Crystallization of lysine and arginine rich proteins
From: Enrico Stura estura {- at -} CEA {- dot -} FR
Date: 2009-11-02