redox buffer novice

Kresten kresten at
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 ?

Pyridine-ethylation and Amino acid analysis? ...


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

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Kresten Lindorff Larsen, Dept. Yeast Genetics

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