Table 1.
Pks13, Mas and PpsA constructs and their global and hydrodynamic properties
| Protein a | Pks13 (Holo-Pks13) | Pks13(S1533A) monomer (C16-Pks13(S1533A) dimer) | fACP1‑KS‑AT | fKS‑AT | fKS | fAT | AT52 | fACP2‑TE | fTE | Mas monomer (Dimer) | PpsA |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sequence boundaries | 1–1733 | 1–1733 | 1–1063 | 119–1071 | 119–575 | 591–1046 | 576–1063 | 1154–1733 | 1437–1725 | 1–2111 | 1–1876 |
| MWth (kDa), Number of residues | 188.0, 1746 | 188.0, 1746 (376.0, 3492) | 115.8, 1084 | 104.2, 974 | 50.5, 478 | 49.2, 479 | 55.1, 510 | 63.6, 583 | 32.8, 295 | 225.9, 2124 (451.8, 4248) | 200.4, 1889 |
| [NaCl] (mM) | 50 | 50 | 50 | 50 | 50 | 300 | 300 | 50 | 50 | 50 | 500 |
| pH | 8.0 | 8.0 | 8.5 | 8.0 | 8.0 | 8.0 | 8.0 | 7.5 | 8.0 | 8.0 | 8.0 |
| 10% glycérol | 2 mM EDTA | ||||||||||
| 10% glycérol | |||||||||||
| DLSb | |||||||||||
| Rh (nm) | 7.3 (7.9) | 6.9 (8.7) | 4.2 | 4.5 | 3.4 | 3.0 | 3.7 | 3.4 | 2.4 | 6.5 (–) | 8.1 |
| Pd (%) | 17 (25) | 26 (35) | 19 | 23 | 25 | 20 | 15 | 27 | 19 | 13 (–) | 18 |
| SAXSc | |||||||||||
| Rg (nm) | 7.4 ± 0.1/7.7 (7.4 ± 0.1/7.6) | 8.2 ± 0.4/7.9 (10.3 ± 0.1/10.7± 0.1) | 3.8 ± 0.1/4.0 | 3.8/3.8 | 2.8 ± 0.1/2.9 | 2.8 ± 0.0/2.7 | 3.4 /3.5 | 3.1/3.4 | 1.9 ± 0.1/2.0 | 5.7/5.9 (8.0 ± 0.2/8.4) | 6.4 ± 0.3/ 6.5 |
| MW (kDa) | 210/214 (248/255) | −/− | 105/105 | 110/110 | −/− | −/− | 44/44 | 46/47 | 26/27 | −/− | −/− |
| −/− | −/− | ||||||||||
| MWind (kDa) | 214 ± 26/190 (216 ± 22/179) | 227 ± 30/227 (431 ± 36/290) | 116 ± 7/125 | 102 ± 2/115 | 54 ± 7/58 | 54 ± 6/60 | 58 ± 3/64 | 59 ± 1/63 | 27 ± 3/30 | 221 ± 23/225 (334 ± 99 /343 ) | 182 ± 16/217 |
| Dmax (nm) | 24.8 (24.8) | 25.0 (36.0) | 12.7 | 13.0 | 10.0 | 8.0 | 12.0 | 12.0 | 5.5 | 21.0 (29.0) | 20.2 |
| Oligom. state | M (M) | M (D) | M | M | M | M | M | M | M | M (D) | M |
| tRg (nm) d | 3.8/4.9/13.9 | 3.8/4.9/13.9 | 3.1/4.1/ 10.9 | 3.0/3.9/10.3 | 2.3/3.0/7.2 | 2.3/3.0/7.3 | 2.3/3.0/7.3 | 2.5/3.2/8.0 | 1.9/2.5/5.6 | 4.0/5.3/13.9 | 3.9/5.0/14.4 |
| χ DAMMIN/ GASBOR/ | |||||||||||
| Comparison with HR model | 1.36/2.69/3.11 (−/−/−) | 1.16/−/− (1.16/−/−) | 1.80/2.80/1.02 | 1.57/1.83/1.36 | 1.28/1.10/1.11 | 2.91/2.72/1.14 | 2.12/2.68/3.10 | 1.86/4.32/1.97 | 3.41/2.93/1.67 | 1.44/1.54/− | 1.32/1.56/− |
a Theoretical molecular weight and number of residues given for the tagged proteins in the case of Pks13, holo-Pks13, C16-Pks13(S1533A), Mas, PpsA, fACP1‑KS‑AT, fKS‑AT, fKS, fAT, AT52 and after removal of the His6-tag for fACP2‑TE and fTE
b Hydrodynamic radii (Rh) and percentage of polydispersity obtained from dynamic light scattering at 20 °C. Histograms of Rh can be found in Additional file 1: Table S1
cRg, gyration radii based on Guinier plot/distance distribution function. MW, molecular weight based on I0 from Guinier plot/I0 from distance distribution function p(r). MWind, molecular weight based on the concentration-independent methods by Rambo and Tainer/SAXSMoW calculator. Dmax maximum dimension. Oligomeric state (monomer, M; partial dimerization, D) obtained from SAXS (at 12–15 °C). For data arising from online HPLC measurements, only the concentration-independent evaluation of the molecular weight is given. Error values are given except when their mathematical rounding gives 0
d Theoretical radii of gyration calculated for a monomeric globular/dimeric globular/unfolded protein