Table 3 In vitro reduction of NOHA by recombinantly expressed hmARC isoforms

A complete incubation mixture consisted of 10 μg of mARC1 or mARC2, 200 pmol of cyt b5, 20 pmol of cyt b5 reductase, 1 mM NADH and 0.5 mM substrate in 150 μl of 20 mM Mes buffer, pH 6.0. Results are means±S.D. for three different incubations each analysed twice via HPLC.

mARC isoformCompositionSpecific activity (nmol·min−1·mg−1 of total protein)
NOHA reductionNHAM reduction
Recombinant hmARC1Complete39.1±1.420.6±0.9
Without cyt b511.5±2.78.9±0.2
Without cyt b5 reductase1.6±0.30.7±0.2
Without hmARC11.2±1.10.5±0.5
Only hmARC11.8±0.41.6±0.3
Without NADH0.5±0.50.1±0.1
Recombinant hmARC2Complete56.8±5.07.7±0.4
Without cyt b51.7±1.70.2±0.2
Without cyt b5 reductase1.0±0.40.7±0.3
Without hmARC20.1±0.10.2±0.2
Only hmARC20.0±0.00.3±0.3
Without NADH0.1±0.10.1±0.1