TABLE 1.
Copper binding stoichiometries of Sco1 mutants
Recombinant soluble His-Sco1 mutant proteins were purified from E. coli cultured in medium containing supplemental copper. Copper per protein stoichiometries were measured from protein samples as purified, following concentration, as well as 12 h dialysis. Cu(II) binding was assessed by comparing A360 of mutants to A360 of wild-type His-Sco1 at the same protein concentrations.
|
Cu/protein |
WT | Y219D | R220D | V221D | Y222D |
|---|---|---|---|---|---|
| Sco1 | Sco1 | Sco1 | Sco1 | Sco1 | |
| Elution | 1.0 ± 0.1 | 1.0 ± 0.1 | 0.9 ± 0.1 | 0.9 ± 0.1 | 0.9 ± 0.1 |
| Concentration | 1.0 ± 0.1 | 1.1 ± 0.1 | 0.9 ± 0.1 | 0.8 ± 0.1 | 0.8 ± 0.1 |
| Post-dialysis | 0.9 ± 0.1 | 0.9 ± 0.1 | 0.8 ± 0.1 | 0.7 ± 0.1 | 0.7 ± 0.1 |
| ε Cu(II)/WT ε Cu(II) | 1.0 | 1.0 | 0.9 | 0.8 | 1.1 |