TABLE 4

V_max and K_m values of the reduction of benzamidoxime by mARC-1 WT and multiple variants

Data are the means ± S.D.

mARC Protein^{^a}	Protein Lots	V_max^{^b}	K_m^{^b}
	n	nmol BA/min per mg protein	mM BAO
mARC-1	2	106 ± 25	0.2 ± 0.0
Single variant	1	82 ± 1	0.2 ± 0.0
Double variant	1	110 ± 1	0.3 ± 0.0
Triple variant	1	62 ± 2	0.5 ± 0.0
Quadruple variant	1	52 ± 1	0.4 ± 0.0
Quintuple variant	1	41 ± 5	0.4 ± 0.2
Sextuple variant	1	1 ± 0	0.7 ± 0.2

BA, benzamidine; BAO, benzamidoxime.
↵^a Amino acid substitutions A165T, M187K, V96L, C246S, D247H, and M268I were introduced successively in mARC-1 WT.
↵^b Each mARC protein was incubated in duplicate and measured twice by HPLC. Kinetic parameters were determined as described in Materials and Methods.