Table 1.
VieB specifically interacts with VieS-C
| Protein Combination | Native Oligomeric State | Hetero-association stoichiometry | K D (μM) | |
|---|---|---|---|---|
| VieS-C + VieA-His6 | VieS-C | VieA-His 6 | 1 dimer : 1 dimer | 1.38 ± 0.35 |
| Dimer (MW = 150 kDa) | Dimer (MW = 130 KDa) | |||
| VieS-C + VieB | VieS-C | VieB | 1 dimer : 1 monomer | 0.467 ± 0.054 |
| Dimer (MW = 150 kDa) | Monomer (MW = 64 kDa) | |||
| VieS-C + VieB D62E | VieS-C | VieB D62E | 1 dimer : 1 monomer | 0.197 ± 0.061 |
| Dimer (MW = 150 kDa) | Monomer (MW = 64 kDa) | |||
| VieA-His6 + VieB | VieA-His 6 | VieB | N/A | N/A |
| Dimer (MW = 130 kDa) | Monomer (MW = 64 kDa) | |||
Characterization of VieS-C, VieA-His6, wild-type VieB and the VieB D62E point mutant (self-association) were determined by Size-Exclusion Chromatography Multi-angle Light Scattering. To determine the protein-protein interactions of various VieSAB protein combinations, hetero-association interaction kinetics were determined over a range of protein concentrations by Composition-gradient Multi-angle Light Scattering. Data for the hetero-association stoichiometry are represented in monomer units. These data represent the average and ± SD of three independently purified replicates.