| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
0.1% |
100 |
β |
76 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
2% |
100 |
β |
23 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
1% |
100 |
β |
41 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
0.1% |
100 |
β |
62 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
0.1% |
100 |
β |
69 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
2% |
100 |
β |
5 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
1% |
100 |
β |
42 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
1% |
100 |
β |
20 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
0.1% |
100 |
β |
59 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
2% |
100 |
β |
22 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
2% |
100 |
β |
18 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
0.1% |
100 |
β |
47 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
1% |
100 |
β |
49 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
0.1% |
100 |
β |
79 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
1% |
100 |
β |
14 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
1% |
100 |
β |
66 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
2% |
100 |
β |
2 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| WT |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
2% |
100 |
β |
62 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
2% |
100 |
2 |
4 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
2% |
100 |
2 |
2 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
2% |
100 |
23 |
18 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
2% |
100 |
23 |
22 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
0.1% |
100 |
69 |
70 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
1% |
100 |
14 |
4 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
0.1% |
100 |
69 |
94 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
1% |
100 |
14 |
10 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
1% |
100 |
42 |
43 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
1% |
100 |
42 |
31 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
2% |
100 |
22 |
27 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
0.1% |
β |
47 |
64 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetone |
2% |
100 |
22 |
10 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Isopropanol |
0.1% |
100 |
47 |
50 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
2% |
100 |
18 |
20 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
0.1% |
100 |
76 |
84 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
0.1% |
100 |
76 |
99 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
1% |
100 |
66 |
62 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
1% |
100 |
66 |
61 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
2% |
100 |
62 |
44 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Acetonitrile |
2% |
100 |
62 |
42 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
0.1% |
100 |
79 |
88 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
0.1% |
100 |
79 |
91 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
1% |
100 |
49 |
47 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
1% |
100 |
49 |
20 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
1% |
100 |
41 |
40 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Methanol |
2% |
100 |
18 |
4 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
0.1% |
100 |
59 |
54 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
0.1% |
100 |
59 |
57 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
1% |
100 |
20 |
15 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
1% |
100 |
20 |
3 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
2% |
100 |
5 |
2 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Ethanol |
2% |
100 |
5 |
3 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
0.1% |
100 |
62 |
49 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| I359L |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
0.1% |
100 |
62 |
78 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |
| R144C |
Activity - Classical |
Activity measured by HPLC-coupled mass spectrometry in the presence of organic solvent |
Dimethyl Sulfoxide (DMSO) |
1% |
100 |
41 |
34 |
% |
100 mM potassium phosphate buffer |
7.4 |
37Β°C |
4 Β΅M (S)-Warfarin |
(S)-7-Hydroxywarfarin |
1 mM NADPH |
β |
Classical aqueous control (in %) |