TABLE 1.
Enzyme activities of the membrane-bound NDH-1 of E. coli wild-type and various NuoI mutants
| Sample | dNADH-O2a | dNADH-DBa | dNADH-K3Fe(CN)6b |
|---|---|---|---|
| WT | 766 ± 35 (100%) | 786 ± 35 (100%) | 1815 ± 38 (100%) |
| NuoIKO | 16 ± 5 (2%) | 19 ± 5 (2%) | 301 ± 35 (17%) |
| KO-rev | 740 ± 76 (97%) | 1003 ± 59 (128%) | 1920 ± 72 (106%) |
| C60A | 10 ± 8 (1%) | 19 ± 1 (2%) | 312 ± 12 (17%) |
| C60S | 39 ± 4 (5%) | 57 ± 5 (7%) | 561 ± 52 (31%) |
| C60H | 12 ± 3 (2%) | NDc | 365 ± 26 (20%) |
| C63A | 18 ± 2 (2%) | 17 ± 4 (2%) | 409 ± 17 (23%) |
| C63S | 153 ± 32 (20%) | 160 ± 15 (20%) | 905 ± 67 (50%) |
| C66S | 13 ± 2 (2%) | 16 ± 3 (2%) | 323 ± 11 (18%) |
| C66H | 15 ± 1 (2%) | 7 ± 2 (1%) | 352 ± 16 (19%) |
| C70S | 28 ± 4 (4%) | 31 ± 1 (4%) | 343 ± 19 (19%) |
| C70H | 4 ± 1 (1%) | 12 ± 1 (2%) | 265 ± 8 (15%) |
| C99S | 15 ± 2 (2%) | 18 ± 1 (2%) | 308 ± 4 (17%) |
| C99H | 8 ± 1 (1%) | ND | 268 ± 9 (15%) |
| C102S | 14 ± 1 (2%) | 38 ± 2 (5%) | 273 ± 9 (15%) |
| C102H | 9 ± 1 (1%) | 15 ± 1 (2%) | 307 ± 27 (17%) |
| C105S | 11 ± 3 (1%) | 12 ± 1 (2%) | 312 ± 14 (17%) |
| C109S | 16 ± 1 (2%) | 29 ± 8 (4%) | 380 ± 48 (21%) |
| T30A | 470 ± 50 (61%) | 453 ± 43 (58%) | 1169 ± 27 (64%) |
| P34A | 499 ± 20 (65%) | 450 ± 16 (57%) | 1167 ± 79 (64%) |
| P42A | 468 ± 27 (61%) | 510 ± 30 (65%) | 1964 ± 78 (108%) |
| R43A | 350 ± 1 (46%) | 418 ± 13 (53%) | 1520 ± 18 (84%) |
| Y44A | 920 ± 3 (120%) | ND | 2520 ± 12 (139%) |
| G46A | 598 ± 6 (78%) | ND | 1437 ± 33 (79%) |
| R52A | 571 ± 21 (75%) | 504 ± 20 (64%) | 1532 ± 33 (84%) |
| P54A | 563 ± 3 (73%) | 623 ± 42 (79%) | 1717 ± 20 (95%) |
| G56A | 362 ± 11 (47%) | 424 ± 18 (54%) | 1701 ± 46 (94%) |
| E58A | 651 ± 6 (85%) | 503 ± 19 (64%) | 1789 ± 8 (99%) |
| V61A | 238 ± 27 (31%) | 257 ± 18 (33%) | 1225 ± 31 (67%) |
| L65A | 511 ± 49 (67%) | 568 ± 97 (72%) | 1345 ± 17 (74%) |
| P71A | 347 ± 73 (45%) | 437 ± 42 (56%) | 1364 ± 20 (75%) |
| I75A | 218 ± 58 (28%) | 318 ± 83 (40%) | 1191 ± 41 (66%) |
| G85A | 885 ± 57 (116%) | 688 ± 42 (88%) | 1929 ± 23 (106%) |
| F92A | 620 ± 42 (81%) | 599 ± 58 (76%) | 1436 ± 30 (79%) |
| R93A | 777 ± 67 (101%) | 719 ± 55 (92%) | 1837 ± 50 (101%) |
| I94A | 704 ± 3 (92%) | 777 ± 11 (99%) | 1954 ± 26 (108%) |
| I94G | 715 ± 3 (93%) | 935 ± 28 (119%) | 1678 ± 32 (92%) |
| R98A | 151 ± 20 (9%) | 141 ± 18 (18%) | 953 ± 44 (53%) |
| I100A | 649 ± 4 (85%) | 676 ± 14 (86%) | 1827 ± 78 (101%) |
| I100G | 471 ± 10 (61%) | 665 ± 8 (85%) | 1565 ± 5 (86%) |
| G103A | 766 ± 4 (100%) | 631 ± 69 (80%) | 1604 ± 36 (88%) |
| E107A | 441 ± 99 (58%) | 556 ± 86 (71%) | 1211 ± 38 (67%) |
| P110A | 71 ± 8 (9%) | 151 ± 8 (19%) | 1421 ± 22 (78%) |
| I114A | 715 ± 31 (107%) | 954 ± 29 (121%) | 1710 ± 60 (94%) |
| E121A | 569 ± 3 (74%) | 671 ± 32 (85%) | 1798 ± 26 (99%) |
| Y132A | 645 ± 32 (84%) | 829 ± 7 (105%) | 2233 ± 21 (123%) |
a Activity is shown in nanomoles of dNADH/mg of protein/min.
b Activity is shown in nanomoles of K3Fe(CN)6/mg of protein/min.
c ND means not determined.