Table 3.
apoMS treatment | MS activity, nmol/min |
---|---|
MeCbl | |
w/hMSRI-H and NADPH | 0.83 |
w/hMSRI-H | 0.62 |
w/NADPH | 0.39 |
w/o hMSRI-H and NADPH | 0.35 |
aqCbl | |
w/hMSRI-H and NADPH | 0.24 |
w/hMSRI-H | 0.11 |
w/NADPH | 0.02 |
w/o hMSRI-H and NADPH | 0.01 |
No Cbl | |
w/hMSRI-H and NADPH | 0.06 |
w/o hMSRI-H and NADPH | <0.01 |
A crude extract from Sf9 cells infected with recombinant baculovirus was used as the source of human apoMS. A mixture (total volume was 200 μl) that contained 150 μl of crude extract (2.2 mg/ml protein) and the components indicated (w/, with; w/o, without) was preincubated for 15 min at 37°C prior to measurement of MS activity by the aqCbl/DTT assay. The concentrations of the components were 50 μM MeCbl or aqCbl, 200 nM hMSRI-H, and 200 μM NADPH.