Table 3.
Binding of ligands from crude reaction mixtures (CRMs) in Hsp90 crystals is not related to affinity.
| CRM no | Solubility of pure (µM) | Result soak of pure | Result soak of CRM | koff CRM (s−1) | Yield (%) | HAC | nRot | KD (SPR) (µM) of pure | KD (ITC) (µM) of pure |
|---|---|---|---|---|---|---|---|---|---|
| 2 | 400 | Bound | PR | 0.1 | 8 | 22 | 1 | 1.0 | 0.65 |
| 11 | 24 | Native | SM1 | 0.23 | 34 | 30 | 4 | 17 | ND |
| 15 | 440 | Bound | SM1 | 0.06 | 1 | 20 | 3 | 0.07 | 0.36 |
| 63 | 460 | Bound | SM2 | 0.012 | 9 | 33 | 4 | 0.04 | 0.07 |
| 57 | 320 | Bound | SM2 | 0.03 | 39 | 32 | 3 | 0.71 | 0.26 |
| 68 | 350 | Bound | SM2 | 0.12 | 9 | 29 | 4 | 3.0 | ND |
| 72 | 300 | Bound | SM2 | 0.12 | 8 | 30 | 6 | 1.7 | 3.3 |
| 75 | 7 | Bound | SM2 | 0.013 | 3 | 30 | 3 | 0.019 | ND |
| 83 | 370 | Bound | SM2 | 0.05 | 8 | 31 | 6 | 0.36 | 0.29 |
| SM1 | >1 | 2000 | |||||||
| SM2 | 0.2 | 2 |
The results for measuring solubility at pH 7.4 and affinity and manual crystal structure determination for selected CRMs and associated purified products. Solubility, SPR and ITC measurements were made at 25 °C. SPR measurements are an average of at least two determinations; ITC and solubility are a single measurement. ND in ITC was due to lack of heat observed.