redox buffer novice

Kresten kresten at my-deja.com
Wed Jun 21 07:10:09 EST 2000


>
> I am studying the redox properties of my enzyme using various redox
> buffers (GSH/GSSG; cysteine:cystine; DTTred/DTToxid).  Most of the
> protocols that have described in the literature for these buffers
> don't
> describe whether these buffers were deoxygenated prior to use; or
> whether any precautions were taken to prevent oxidation during the
> assay.  Are there any protocols for redox buffers that take the effect
> of oxygen into account ?

I usually add a bit of EDTA (preventing metal-ion catalyzed oxidation)
and purge extensively with Argon.

>
> One more question: the active site cysteines of may enzyme may become
> oxidised to sulfinic or sulfenic acids ... how could I test this ?

Ellman?
Sequencing?
IEF?
Pyridine-ethylation and Amino acid analysis? ...

HTH
Kresten

>
> Thank you for your time,
>
> Che Pillay
> Dept. of  Biochemsitry
> University of Natal
> South Africa
> pillayche at nu.ac.za
>
> ---
>

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Kresten Lindorff Larsen, Dept. Yeast Genetics


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