Table 3.
Results of sedimentation velocity analytical centrifugation analysis (SV-AUC) of N130 in the absence or presence of increasing concentrations of the rpL5 peptide.
| rpL5:N130 | mg/mla | s20 (Svedberg)b | s20,w (Svedberg)c | MW (kDa)d | f/f0 e |
|---|---|---|---|---|---|
| 0:1 | 2.76 | 4.63 (98.7%) | 4.82 | 88.9 | 1.53 |
| 7.90 (1.20%) | 8.22 | 198.0 | 1.53 | ||
| 1:1 | 3.09 | 5.23 (96.3%) | 5.44 | 99.3 | 1.46 |
| 7.63 (3.7%) | 7.94 | 175.0 | 1.46 | ||
| 2:1 (1-300 min) | 3.63 | 1.07 (2.6%) | 1.11 | 6.8 | 1.20 |
| 5.80 (48%) | 6.03 | 86.3 | 1.20 | ||
| 8.11 (23%) | 8.44 | 143.0 | 1.20 | ||
| 12.33 (25%) | 12.82 | 268.0 | 1.20 | ||
| 14-19 (7%) | 14-19 | 300-500 | 1.20 | ||
| 2:1 (100-300 min)f | 3.31 | 5.75 (56%) | 5.98 | 81.1 | 1.16 |
| 5.80 (25%) | 6.03 | 128.0 | 1.16 | ||
| 8.11 (17%) | 8.44 | 182.0 | 1.16 | ||
| 3:1 (1-300 min) | 2.01 | 0.95 (17%) | 0.98 | 7.4 | 1.43 |
| 5.80 (31%) | 6.00 | 112.0 | 1.43 | ||
| 10.32 (32%) | 10.73 | 266.0 | 1.43 | ||
| 14-27 (21%) | 14-27 | 500-850 | 1.43 | ||
| 3:1 (100-300 min)f | 1.29 | 1.18 (4%) | 1.22 | 1.1 | 1.49 |
| 5.73 (41%) | 5.96 | 117.0 | 1.49 | ||
| 7.41 (20%) | 7.71 | 173.0 | 1.49 | ||
| 9.17 (15%) | 9.54 | 238.0 | 1.49 | ||
| 11.33 (15%) | 11.79 | 266.0 | 1.49 |
a Total concentration in mg/ml.
b Sedimentation coefficient taken from the ordinate maximum of each peak in the best-fit c(s) distribution at 20 °C with percentage protein amount in parenthesis. Sedimentation coefficient (s-value) is a measure of the size and shape of a protein in a solution with a specific density and viscosity at a specific temperature.
c Standard sedimentation coefficient (s20,w-value) in water at 20 °C.
d Molar mass values (MW) taken from the c(s) distribution that was transformed to the c(M) distribution. The theoretical molar mass is in parenthesis.
eBest-fit weight-average frictional ratio values (f/f0)w taken from the c(s) distribution.
f Fit of the data in the 100-300 min sedimentation time frame, for the analysis of the lower MW rpL5:N130 complex intermediates. The MW cutoff of ~ 300 kDa corresponds to a tetramer of N130 pentamers.