The effect of 17O-labeled
H2O2 on DEPMPO/⋅OH formation.
Incubation mixtures contained H2O2 (5 mM),
DEPMPO (20 mM), and DTPA (100 μM) in a bicarbonate/CO2
buffer. The reaction was initiated with either Fe(II)-EDTA (100 μM)
or 100 μg of SOD or SOD mutants (G93A and A4V) as shown. Line
positions from 17O and 16O couplings are shown
by • and ○, respectively. The dotted lines show a
computer simulation of ESR spectra using the parameters
aN = 14.0 G, aH = 13.2 G,
aP = 47.0 G for
DEPMPO/⋅16OH and aN = 14.0
G, aH = 13.1 G, aP = 47.0
G, and a17O = 4.1 G for
DEPMPO/⋅17OH).