Figure 5
Assembly of RsbV–RsbW in solution. AF4-MALS profiles of (a) RsbV–RsbW, (b) RsbV1–104–RsbW5–145, (c) RsbW–SigB, (d) RsbV and (e) RsbW. The thick lines represent the molar masses of the proteins. All AF4-MALS profiles exhibit distinct single peaks, indicating that the protein solution does not contain any other oligomers or aggregates. The molecular masses of RsbV–RsbW and RsbV1–104–RsbW5–145 were calculated to be 114.5 and 97.0 kDa, respectively, which are close to the theoretical molecular masses of the hetero-octamers RsbV–RsbW (120.8 kDa) and RsbV1–104–RsbW5–145 (111.2 kDa). The molar masses of RsbV and RsbW were 12.3 and 66.7 kDa, indicating that RsbV and RsbW exist as monomers and tetramers in solution, respectively. The molar mass of RsbW–SigB was 115.8 kDa, which is close to that of an assembly of four RsbW and two SigB molecules. |