LC-MS analysis of the transformations of L-hArg deuterium isotopologs by VioC,
and interpretation of the results in terms of mechanistic pathways,
2H-KIE
(kH/kD) values, and
impacts on partition ratios. A, All-protium
L-hArg. B, C3-labeled L-Arg
(red), C4-labeled L-Arg
(blue), and uniformly labeled L-Arg
(purple). The red values are partition
ratios for (i) the initial HAT to the ferryl complex from C3 or C4
(upper in
A), (ii) the fate of the C3 radical, rebound or
second HAT (middle in
A), and (iii) overall products
(lower in A
and B). The red numbers in parentheses are the
standard deviations of the values from three independent trials. The
2H-KIE values that rationalize the impacts of deuterium
substitution on the partition ratios are shown in gray and black (in
A). The arrows and dashes (in
B) illustrate the qualitative impacts of
the 2H-KIEs on the partitions between HAT from C3 or C4 to the ferryl
complex (gray), between the two pathways for decay
of the C3 radical (black), and among the three
possible products (gray and
black). Details of the analytical
procedure are provided in the Supporting Information. The values resulting from product detection
by A260 (from the Fmoc group) are compared in Table S3.