In article <4b4jj7$nc3 at milo.mcs.anl.gov> Mark Fuller <mark_fuller at qmgate.anl.gov> writes:
>Does anyone have some general guidelines for inhibiting electron
>transport? I plan to work with rotenone antimycin A and
>2-N-heptyl-4-hydroxyquinolane-N-oxide and pure cultures of bacteria.
You need then to recognise that not all bugs (and actually rather few)
are inhibited by 'classical' (mitochondrial) electron transport inhibitors.
The ones you mention work at ca 1 micromolar or 1 nmol/mg if the bugs
are 1 mg/ml (correct usage, as most probes are bound). People tend
to disbelieve that you are being specific if you whack the concentration
up much beyond that.
>What is the recommended concentrations for these compounds? Are there
>any special culture conditions that should be maintained to allow these
>compounds to work?
Don't wait too long; bugs will produce e.g. rotenone-insenitive
ET chains in culture if rotenone is present, or at least Paracoccus
does (Phillips, M.K. & Kell, D.B. (1982) A novel inhibitor of NADH
dehydrogenase in Paracoccus denitrificans. FEBS Lett. 140, 248- 250).
>>Any advise, especially refererences, would be greatly appreciated.
>>Mark Fuller, Ph. D.
>Environmental Research Division
>Argonne National Laboratory
>Argonne, IL 60439-4843
>>phone: (708) 252-1795
>FAX: (708) 252-8895
>email: mark_fuller at qmgate.anl.gov>>Best of luck,
Douglas.
