Table 1.
Functionality and yield of purified Ste2p-rho from 1 L culture.
Receptor and Strain | Growth arrest assay (%) | Kd (nM) | Bmax or Yield |
---|---|---|---|
WT (C59S, C252S) in LM102 | 100 ± 12 | 8.9 ± 0.6 | 75,300 ± 6800 per cell |
Ste2p-rho in LM102 | 104 ± 7 | 12.7 ± 1.2 | 36,800 ± 3900 per cell |
Purified Ste2p-rho from BJS21 | Not Applicable | 27.8 ± 3 | 120 μg/100 mg membrane (26 pmole/mg)
39 ± 7 % active receptora |
Active population of the purified receptor was calculated from the Bmax value in saturation binding assays. Specific activity of [3H]α-factor was 16 Ci/mmol and the average Bmax value of four separate binding assays was 6,000 cpm/40 ng receptor.