Table 2.
Copper content of cgAUS1 wild type and mutants determined by atomic absorption spectroscopy (AAS). Protein concentration in mg/ml of cgAUS1 was determined after purification by UV at 280 nm; molecular weight (MW) and extinction coefficient for the protein concentration in μM was calculated by ProtParam [50]; copper concentration was determined by AAS; Copper/cgAUS1 is the determined ratio of copper per molecule AUS.
| Sample | Protein [mg/ml] | MW [g/mole] | Protein [μM] | Copper [μM] | Copper/cgAUS1 |
|---|---|---|---|---|---|
| AUS1 w/t | 0.63 | 85 761 | 7.39 | 15.42 | 2.1 |
| AUS1 H93A | 0.37 | 85 694 | 4.36 | 4.56 | 1.0 |
| AUS1 H116A | 1.09 | 85 694 | 12.67 | 14.16 | 1.1 |
| AUS1 H125A | 0.74 | 85 694 | 8.65 | 8.97 | 1.0 |
| AUS1 F273A | 0.27 | 85 684 | 3.17 | 7.40 | 2.3 |
| AUS1 C97A | 0.06 | 85 728 | 0.67 | 1.38 | 2.0 |