Deamino nadh

Antibiotics, 5-aminolevulinic acid hydrochloride (5-ALA), ß-mercaptoethanol, ß-NADH, bovine xanthine oxidase, casein acid hydrolysate, cytochrome c from equine heart, deamino-NADH, desferoxamine mesylate (DFO), diethylenetriamine pentaacetic acid (DTPA), 2,2’-dipyridyl (DIP), ethyl acetate, ferric chloride, ferrous ammonium sulfate, 30% hydrogen peroxide, 8-hydroxyquinoline-5-sulphonic acid, E. coli manganese-containing superoxide dismutase, manganese (II) chloride tetrahydrate, 2-[N-morpholino]ethanesulfonic acid (MES), o-dianisidine dihydrochloride, o-nitrophenyl-ß-D-galactopyranoside (ONPG), potassium ferricyanide, potassium cyanide, tricine, protoporphyrin IX, and xanthine were purchased from Sigma. Ethylenediamine tetraacetic acid (EDTA), guanidine hydrochloride, hydrochloric acid, and 3-(N-morpholino) propane-sulfonic acid (MOPS) were purchased from Fisher Scientific; Coomassie protein assay reagent and albumin standard, from Thermo Scientific; sodium dithionite, from Fluka; and glacial acetic acid, from J.T.Baker.

Oxygen uptake rates were measured polarographically with a Clark-type oxygen electrode (Rank Brothers, Cambridge, UK) at 25 °C in 1 mL of air-saturated medium containing 0.3 M sucrose, 10 mM KH₂PO₄, 10 mM TES and 10 mM MgCl₂ (pH 6.8). NADH:ubiquinone oxidoreductase activity was measured for 40 µg of mitochondria which were freeze–thawed three times in the presence of 1 mM deamino-NADH (Sigma-Aldrich, St. Louis, USA). Percentage FSL0260 resistance was calculated using the formula in Eq. (1): $$\%\mathrm{FSL0260}\mathrm{resistance}=\left\{\right[\mathrm{Rate}+\mathrm{FSL0260}]/[\mathrm{Rate}-\mathrm{FSL}0260\left]\right\}\times 100\%$$ where Rate is the rate of oxygen consumption (nmol oxygen min⁻¹ mg⁻¹ of protein).