TABLE 1.
In vitro assay of FACR and FALDR activities in maFACR variants.
| maFACR varianta |
FACR specific activityb |
FALDR specific activityc |
||
| (nmol NTB min–1 mg–1 protein) | (nmol NTB min–1 nmol–1 protein) | (nmol NADP+ min–1 mg–1 protein) | (nmol NADP+ min–1 nmol–1 protein) | |
| maFACRWT | 63.2 ± 1.5 | 7.3 ± 0.2 | 7789.3 ± 43.8 | 905.3 ± 5.1 |
| maFACRS126D | 60.9 ± 8.5 | 7.1 ± 1.0 | ND | ND |
| maFACRK156A | 73.2 ± 2.3 | 8.5 ± 0.3 | ND | ND |
| maFACRY152F | 64.1 ± 0.9 | 7.4 ± 0.1 | ND | ND |
| maFACRSYK | 70.2 ± 0.8 | 8.2 ± 0.1 | ND | ND |
| maFACRCter | 3.0 ± 0.2 | 0.23 ± 0.02 | ND | ND |
| maFACRS515A | ND | ND | 2,594.0 ± 285.0 | 301.5 ± 33.1 |
| maFACRK527A | ND | ND | 2,689.0 ± 73.1 | 312.5 ± 8.5 |
| maFACRY532F | ND | ND | 2,440.6 ± 263.1 | 283.7 ± 30.6 |
| maFACRNter | ND | ND | 12,875.0 ± 291.2 | 1,104.2 ± 25.0 |
amaFACRWT is the wild-type enzyme. maFACRSYK has the mutations S126D, Y152F, and K156A. maFACRNter and maFACRCter are the N- and C-terminal domains, respectively. Otherwise, maFACRX refers to a single-site mutant, whereby the mutation X is S126D, Y152F, K156A, S515A, Y527F, or K532A.bPalmitoyl-CoA was the substrate used.cDecanal was the substrate used.ND, not detected; FACR, acyl-CoA reductase; FALDR, fatty aldehyde reductase.