Table 1.
Metal content and molecular masses of HvJAMM1 wild-type (WT) and variant proteins.
| HvJAMM1 | Metal content (mol/mol protein)1 | Average Mass (Da)2 | Mass Spec | ||
|---|---|---|---|---|---|
| Zinc | Nickel | Theoretical | Observed | ||
| WT | 0.84 (0.12) | 0.24 (0.018) | 16,771.17 | 16,771.40 | ESI-TOF |
| 16,771.63 | MALDI-TOF | ||||
| E31D | n.d. | n.d. | 16,757.14 | 16,757.21 | ESI-TOF |
| H88N | 0.01 | 0.07 | 16,748.13 | 16,749.74 | MALDI-TOF |
| H90Q | 0.04 | 0.04 | 16,762.16 | 16,762.88 | MALDI-TOF |
| D94N | n.d. | n.d. | 16,770.19 | 16,770.03 | ESI-TOF |
| S98A | 0.37 | 0.13 | 16,775.17 | 16,775.46 | MALDI-TOF |
| D101E | 1.1 | 0.34 | 16,785.20 | 16,785.75 | MALDI-TOF |
| C115S | n.d. | n.d. | 16,755.11 | 16,755.11 | ESI-TOF |
1
Metal content determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES) of HisTrap HTP purified proteins. Metal content of HvJAMM1 purified by size exclusion chromatography in parenthesis.
2
Average mass in Da with theoretical mass based on the HvJAMM1 (HVO_2505) polypeptide sequence with addition of an N-terminal poly-histidine linker (His6-tag) (MGSSHHHHHHSSGLVPRGSH-) and amino acid modifications as indicated. Observed masses are based on MALDI-TOF MS and ESI-MS as indicated.
Abbreviations: WT, wild-type; n.d., not determined.