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

- 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: Crystallization of lysine and arginine rich proteins
From: Flip Hoedemaeker flip {- at -} FORMULATRIX {- dot -} COM
Date: 2009-11-02

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

Subject: Re: Crystallization of lysine and arginine rich proteins
From: Simon Bushell srbushellbbs {- at -} GOOGLEMAIL {- dot -} COM
Date: 2009-11-02

http-equiv="Content-Type">

Following on from Stephen's comment. Jena Biosciences sell a kit that
allows you to do it quickly and easily.

check it out here:
http://www.jenabioscience.com/cms/en/1/catalog/529_jbs_methylation_kit.html

We have had success in our lab using lysine methylation to crystallise
an otherwise recalcitrant protein.

Good luck

Simon

Stephen Graham wrote:
cite="mid:cdd145f50911020556r7bfb367breb18bc1824029890@mail.gmail.com"
type="cite">

Try reductive methylation of the lysines:
Walter, T. S., Meier, C., Assenberg, R., Au, K. F., Ren, J., Verma,
A., Nettleship, J. E., Owens, R. J., Stuart, D. I. & Grimes, J. M.
(2006). Structure, 14, 1617–1622.

Cheers,

Stephen

2009/11/2 Jan Rash <janrsh@googlemail.com>:

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

CCP4bb navigation

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

Previous message:
Subject: Re: Crystallization of lysine and arginine rich proteins
From: Flip Hoedemaeker flip {- at -} FORMULATRIX {- dot -} COM
Date: 2009-11-02

Next 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