Re: [ccp4bb] protein degraded when setting xtal?

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

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

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Subject: Re: protein degraded when setting xtal?
From: Roger Rowlett rrowlett {- at -} MAIL {- dot -} COLGATE {- dot -} EDU
Date: 2008-03-03

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From: Philippe DUMAS p {- dot -} dumas {- at -} IBMC {- dot -} U-STRASBG {- dot -} FR
Date: 2008-03-03

Subject: Re: protein degraded when setting xtal?
From: Kevin Jude kjude {- at -} BERKELEY {- dot -} EDU
Date: 2008-03-03

If your protein is autolyzing, you may need to crystallize it in the
presence of an inhibitor. In the case of trypsin, I have crystallized
in the presence of benzamidine, then removed the inhibitor by dialysis
after crystallization.

kmj

Ngo Duc Tri wrote:
> Dear CCP4 experts,
> I'm working on a kind of cystein peptidase. It shows a little
> degradation after 2 days storing at 4 degree even I tried many kinds of
> protease inhibitors, or glycerol and DTT.
> So I decided to work fast (1 day) then set xtal. After one week I pick
> up some condition and check degradation by SDS-PAGE. It shows heavy
> degradation then I think it's impossible to obtain the crystal if I
> don't find out how to stop the degradation first.
>
> Do you have any idea about my situation? Is it true that I need to stop
> the degradation and how to deal with it?
> Thank you for your advices,
>
> My best regards,
> TriNgo
>
>

CCP4bb navigation

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

Previous message:
Subject: Re: protein degraded when setting xtal?
From: Roger Rowlett rrowlett {- at -} MAIL {- dot -} COLGATE {- dot -} EDU
Date: 2008-03-03

Next message:
Subject: Re: radiation damage question
From: Philippe DUMAS p {- dot -} dumas {- at -} IBMC {- dot -} U-STRASBG {- dot -} FR
Date: 2008-03-03