Table 1.
Expression of wild type and mutant cyt b561 in the bacterial system
| Construct | Total recombinant cyt b561 proteina (nmol/L culture) | Extractable recombinant cyt b561 proteinb (nmol/L culture) | Functional cyt b561c (nmol/L culture) |
|---|---|---|---|
| Wild type | 162 (100%) | 169 (100%) | 134 (100%) |
| N78K | 254 (157%) | 247 (146%) | 227 (169%) |
| H54Q | 1.4 (0.9%) | NDd | NDd |
| H88Q | 41 (25%) | 9.3 (5.5%) | NDd |
| H92Q | 78 (48%) | 69 (41%) | 52 (39%) |
| H109Y | 128 (79%) | 116 (69%) | 117 (87%) |
| H110Q | 120 (74%) | 115 (68%) | 152 (113%) |
| H122Q | 1.7 (1.0%) | NDd | NDd |
| H161Q | 40 (25%) | 13 (7.7%) | NDd |
a
Immunoreactive protein in cell lysate determined by dot blot assay; the percentage of the wild type value is given in parentheses.
b
Immunoreactive protein in detergent extract determined by dot blot assay; the percentage of the wild type value is given in parentheses.
c
Ascorbate-reducible cytochrome in detergent extract calculated from A561(red)-A575(ox) and the difference extinction coefficients determined for the individual purified proteins; the percentage of the wild type value is given in parentheses.
d
Not detected.