. Author manuscript; available in PMC: 2009 Nov 11.

Published in final edited form as: Biochemistry. 2008 Oct 16;47(45):11725–11734. doi: 10.1021/bi801231m

Table 2.

Kinetic parameter estimates for hydroxylation of (S)-warfarin and flurbiprofen.

	(S)-warfarin			Flurbiprofen
	K_mμM	k_catmin⁻¹	k_cat/K_m^bmin⁻¹ μM⁻¹	K_mμM	k_catmin⁻¹	k_cat/K_m^bmin⁻¹ μM⁻¹

WT	4.1 ± 0.3	0.25 ± 0.004	0.06 (100)	22.9 ± 3.7	9.0 ± 0.4	0.4 (100)
F100L	4.5 ± 0.5	0.35 ± 0.01	0.08 (133)	34.8 ± 7.0	12.3 ± 0.7	0.4 (100)
F114L	23.9 ± 4.5^a	0.028 ± 0.002	0.001 (2)	42.8 ± 4.3	4.8 ± 0.1	0.1 (28)
F476L	21.0 ± 2.7^a	0.022 ± 0.001	0.001 (2)	39.2 ± 8.3	4.5 ± 0.2	0.1 (29)
F114W	1.5 ± 0.1	0.078 ± 0.001	0.05 (83)	19.5 ± 3.2	9.2 ± 0.4	0.5 (120)
F476W	3.9 ± 0.6	1.08 ± 0.04	0.3 (500)	104.9 ± 3.9	1.6 ± 0.2	0.02 (5)
F114L/F476L	66.0 ± 12.3^a	0.015 ± 0.001	0.0002 (0.4)	218.6 ± 24.2	3.0 ± 0.1	0.01 (3)
F114W/F476W	2.9 ± 0.3	0.283 ± 0.005	0.1 (167)	237.4 ± 11.8	3.1 ± 0.08	0.01 (3)

Major metabolite formed is 4′-hydroxywarfarin, not 7-hydroxywarfarin. Metabolite formation appears to demonstrate biphasic kinetics, and the values listed are K_m1, k_cat1, and k_cat1/K_m1, respectively.

Bold number in parentheses refers to percent of wild-type k_cat/K_m