Table 1.
Overall biophysical parameters of yCAF1. Column labeling: SEC-MALLS (Size-exclusion chromatography - multi-angle laser light scattering); EQ-AUC (equilibrium analytical ultracentrifugation); SV-AUC (sedimentation velocity analytical ultracentrifugation); SAXS (small angle X-ray scattering); Native MS (native mass spectrometry); MMSLS (Molar masses determined by SEC-MALLS); MMAUC (Molar masses determined by equilibrium analytical ultracentrifugation); s020,w (Sedimentation coefficient determined by velocity analytical ultracentrifugation); sth (computed sedimentation coefficient derived from SAXS envelopes); Rg (radius of gyration); MMSAXS (molar masses determined by SAXS); Dmax (maximum dimension); Vp (excluded particle Volume); MMMS (molar masses determined by native MS; MMth (theoretical molar mass calculated).
The errors reported for SEC-MALLS are the residual standard deviations of the observed data from the fitted values calculated using Astra. The errors of the AUC experiments are derived from the standard deviations of linear fits of the obtained data points to extrapolate the respective values (MMAUC and s020,w) to zero protein concentration. The errors reported for the parameters derived from SAXS are based on the observed range of results is it possible to obtain, adjusting (within acceptable theoretical limits) the data points used for the calculation and as such represent the confidence range of the parameter. Resolution of the ab inito SAXS models was calculated according to (Tuukkanen et al., 2016). Errors in native MS were determined according to (McKay et al., 2006). CSLS, CSAXS, CMS are the concentrations of samples used for SEC-MALLS, SAXS and native MS respectively. N.D. (not determined)
| Sec-malls | Eq-auc | Sv-auc | SAXS | SAXS | SAXS | SAXS | SAXS | Native MS | |||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample |
MMSLS
kDa (CSLS, µM) |
MMAUC
kDa |
sth
Svedberg |
s020,w
Svedberg |
Rg
nm (CSAXS mg·ml−1) |
MMSAXS
kDa |
Dmax
nm |
Vp
nm3 |
Resolution
Å |
MMMS
kDa (CMS µM) |
MMth
kDa |
| yCAF1 | 172.4 ± 1% (67) |
180 ± 10 | 6.2 | 6.41 ± 0.03 | 6.39 ± 0.22 (11.4) |
175 ± 3 | 26 ± 2 | 307 ± 5 | 57 ± 4 | 174.49 ± 0.30 (2.5) |
174.0 |
| yCAF1-H3-H4 | 198.9 ± 1.1% (50) |
200 ± 11 | 7.1 | 6.84 ± 0.06 | 6.02 ± 0.35 (10) |
203 ± 13 | 25 ± 2 | 355 ± 23 | 54 ± 4 | 201.00 ± 0.01 (7) |
200.7 |
| yCAF1T | 142 ± 1% (15) |
N.D. | N.D. | N.D. | 5.66 ± 0.03 (30.0) |
127 ± 1 | 20 ± 1.2 | 222 ± 2 | 49 ± 4 | N.D. | 146.8 |
| yCAF1T-H3-H4 | 153 ± 1% (15) |
N.D. | N.D. | N.D. | 5.10 ± 0.03 (13.2) |
163 ± 1 | 17.3 ± 1.3 | 285 ± 2 | 52 ± 4 | N.D. | 173.6 |