Table 1.
ACAD9 dehydrogenation activity and effects of FAD and ECSIT
| ACAD9 (no His6) | ACAD9-His6 | R532W- His6 | R518H- His6 | R469W- His6 | E426Q- His6 | Loop12- His6 | VLCAD (N- His6) | |
|---|---|---|---|---|---|---|---|---|
| Activity | 83 ± 5 | 80 ± 5 | 62 ± 5∗ | 95 ± 5 | 89 ± 4 | 8 ± 7 | 40 ± 4∗ | 985 ± 75 |
| Activity +100XFAD | 128 ± 5∗∗ | 132 ± 3 | 133 ± 7 | 130 ± 5 | 132 ± 4 | 8 ± 5 | 142 ± 4 | 995 ± 91 |
| Activity + 5X ECSIT | 8 ± 6 | 8 ± 3 | 11 ± 3 | 10 ± 7 | 9 ± 4 | 7 ± 2 | 10 ± 1 | 940 ± 36 |
Reactions contained 0.1 μM purified ACAD9 or VLCAD protein, 50 μM palmitoyl-CoA, and 200 μM ferricenium hexafluorophosphate (15), in a final volume of 0.5 mL. Activity is expressed as μmol ferrocene produced/min/μmol enzyme (min−1). Values are expressed as mean ± SD (n = 3).
∗, p < 0.007 compared to wildtype ACAD9 in the absence of excess FAD.
∗∗, p < 0.0001 compared to wildtype ACAD9 in the absence of excess FAD. See also Figure S2.