Table 2.
Analysis of thiol oxidation by Ero1p-C100A-C105A intragenic suppressors
| Activity (nmol O2 min-1/nmol Ero1p-c)a
|
|||||
|---|---|---|---|---|---|
| Molecular weightb | Wild-type | C100A C105A | L86S C100A C105A | C100A C105A W120R | |
| 5 mM Dithiothreitol | 152 | 16.1 ± 1.9 (129) | 12.5 ± 2.0 (100) | 23.7 ± 4.4 (190) | 64.3 ± 10.7 (514) |
| 50 mM β-Mercaptoethanol | 154 | 3.6 ± 0.2 (109) | 3.3 ± 0.8 (100) | 4.8 ± 0.3 (145) | 6.3 ± 0.3 (191) |
| 50 mM Cysteine | 240 | 5.8 ± 0.8 (187) | 3.1 ± 0.7 (100) | 5.8 ± 0.2 (187) | 8.2 ± 0.5 (265) |
| 50 mM Glutathione | 613 | 1.7 ± 0.1 (189) | 0.9 ± 0.0 (100) | 1.7 ± 0.2 (189) | 3.8 ± 0.5 (422) |
| 0.5 mM Thioredoxin | 12,626 | 75.0 ± 15.0 (2083) | 3.6 ± 0.1 (100) | 8.5 ± 0.7 (236) | 12.5 ± 1.1 (347) |
a
Activity values were extrapolated from the rate of FAD reduction as described in Materials and Methods. Percentage of activity relative to Ero1p-C100A-C105A is noted in parentheses.
b
Molecular weight of the oxidized form.