c-type cytochromes

nsaunders at molbiol.ox.ac.uk nsaunders at molbiol.ox.ac.uk
Thu Apr 18 01:55:36 EST 1996


Hello,

Now then, what species of Pseudomonas are you interested in and is it 
cytochromes c in general you want to look at or one in particular? Both 
Pseudomonas aeruginosa and Pseudomonas stutzeri produce a wide range of 
cytochromes under various growth conditions.

If you're interested in the broad range of cytochromes, a simple way is to 
prepare a total soluble extract, or a periplasmic extract by spheroplasting, 
load it onto a large ion exchange column (e.g. DEAE cellulose) and run a NaCl 
gradient of say 0-400mM. Cytochromes c will elute at various places as 
coloured bands and this is usually enough purification for a crude spectrum. 
You can clean up fractions of interest by a further column, such as gel 
filtration. 
I can supply further details, if you can be more specific.

Neil Saunders




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