Table 2.
SAXS data-collection parameters.
| Instrument | SEC-SAXS at SWING beamline SOLEIL | |||||||||||
| SEC column | Bio-SEC. 3 (Agilent) | |||||||||||
| q range (Å −1) | 0.0022–0.62 | |||||||||||
| Temperature (°C) | 25 °C | |||||||||||
| Structural parameters | ||||||||||||
| From ATSAS/PRIMUS | From ScÅtter | From DAMMIN | ||||||||||
| Sample Name | Sample Concentration (before SEC) | I(0) (cm −1 /absorbance) Reciprocal Space | R g (Å) Reciprocal Space | I(0) (cm −1 /absorbance) Real Space | R g (Å) Real Space | D max (Å) | Porod Volume estimate a (Å 3) | NSD b | χ 2b | Mw from Porod Volume (V Porod /1.6) (Da) | Mw from SAXSMoW (kDa)c | Mw from sequence (kDa) |
| HHARI-RBR | 51 μM | 0.007 | 31.0 | 0.00707 ± 0.00001 | 31.0 ± 0.1 | 112 | 67241 [qmax = 0.16] | 0.63 ± 0.04 | 1.76 | 42025 | 42.0 | 35.8 |
| HOIP-RBR | 200 μM | 0.088 | 31.4 | 0.08845 ± 0.00005 | 31.49 ± 0.05 | 130 | 66111 [qmax = 0.30] | 0.58 ± 0.04 | 1.14 | 41320 | 42.5 | 43.4 |
| TRIAD3A-RBR | 200 μM | 0.037 | 34.9 | 0.03730 ± 0.00003 | 34.93 ± 0.05 | 133 | 61443 [qmax = 0.21] | 0.63 ± 0.02 | 2.38 | 38401 | 36.3 | 34.4 |
| RNF144A-RBR | 200 μM | 0.029 | 29.7 | 0.02981 ± 0.00003 | 29.81 ± 0.05 | 110 | 44641 [qmax = 0.22] | 0.61 ± 0.03 | 1.49 | 27900 | 26.6 | 29.0 |
| HHARI-RBR/UbcH7~Ub | 48 μM | 0.018 | 35.1 | 0.01839 ± 0.00004 | 35.2 ± 0.1 | 128 | 115384 [qmax = 0.20] | 0.58 ± 0.05 | 1.19 | 72115 | 75.3 | 62.5 |
| HHARI-RBR/UbcH5A~Ub | 46 μM | 0.013 | 34.0 | 0.01331 ± 0.00005 | 34.1 ± 0.2 | 128 | 87228 [qmax = 0.18] | 0.57 ± 0.07 | 2.52 | 54517 | 59.0 | 61.3 |
| HOIP-RBR/UbcH7~Ub | 220 μM | 0.13 | 36.7 | 0.1342 ± 0.0001 | 36.91 ± 0.09 | 138 | 118999 [qmax = 0.18] | 0.64 ± 0.06 | 0.54 | 74374 | 74.4 | 70.1 |
| HOIP-RBR/UbcH5A~Ub | 220 μM | 0.07 | 36.9 | 0.0728 ± 0.0001 | 37.1 ± 0.01 | 140 | 100170 [qmax = 0.18] | 0.58 ± 0.03 | 0.53 | 62606 | 74.4 | 68.9 |
| TRIAD3A/UbcH7~Ub | 200 μM | 0.08 | 37.6 | 0.08176 ± 0.00006 | 37.82 ± 0.06 | 151 | 86802 [qmax = 0.22] | 0.67 ± 0.03 | 2.58 | 54251 | 53.3 | 58.1 |
| RNF144A-RBR/UbcH7~Ub | 200 μM | 0.09 | 31.5 | 0.09292 ± 0.00004 | 31.64 ± 0.03 | 118 | 84561 [qmax = 0.20] | 0.50 ± 0.02 | 1.91 | 52850 | 54.9 | 55.7 |
| Software employed | ||||||||||||
| Primary data reduction | Foxtrot | |||||||||||
| Data processing | ATSAS and Scatter | |||||||||||
| Ab initio analysis | DAMMIF/DAMMIN | |||||||||||
| Validation and averaging | DAMAVER | |||||||||||
| Computation of model intensities | CRYSOL | |||||||||||
| 3D graphic representation | PyMOL | |||||||||||
aThe Porod Volume estimation was carried out on truncated data to the highest value of q reported in square brackets.
bValue derived from best DAMMIF model.