Hi Sam,
Depends on your problem. If the cysteins are alone and exposed and is
not involved in multimeric assemly, adding reducing agent helps in
keeping the them(monomers) from non productive association that hinders
crystallization.
On the other hand if they support multimer formation which is
physiological, then it might be more sane to not use reducing agents.
Another worth a penny try is, if you have homology model, try to see if
the cysteins in the same crystal setting helps to form crystal lattice in
the homology model. If yes, then it maybe useful to try not use the reducing
agents in crystallization. If No, try otherwise.
With this in mind you can expand your trials to get your optimum
reducer concentration.
Just my thoughts; may/maynot be useful !
best
Natesh.
On Fri, 27 Jun 2008, Sam wrote:
> Date: Fri, 27 Jun 2008 14:44:02 +0100
> From: Sam <[log in to unmask]>
> To: [log in to unmask]
> Subject: [ccp4bb] Reducer and crystallization
>
> Dear all,
>
> Can anyone enlighten me the effect of reducer in crystallography?
> I understand that it removes disulphide bond, and prevent protein
> aggregation. But how do we find a balance point and must we remove it before
> screening crystals?
>
> Thanks for answering first.
>
>
> Cheers
> sam
>
-------------------------------------
Failures are more finely etched in our minds than triumphs, and success
is an elusive, if not mythic, goal in our demanding society (Hugh Drummond).
-------------------------------------
Ramanathan Natesh
Wellcome Trust Advanced Training Fellow
School of Crystallography
Birkbeck College
University of London
Malet Street
London WC1E 7HX
Tel: +44 (0)20-7631-6835
Fax: +44 (0)20-7631-6803
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