Table 3.
Total functional yields from TM6* producing two human GPCRs are at least quadruple that from the corresponding wild-type transformants
| Transformants | Duration of culture (h) | Specific growth rate (h-1) | Dry cell weight yield (g g-1 Glc) | Harvest wet cell weight (g) | Total membrane protein yield (mg) | Bmax (pmol mg-1) | Kd (nM) |
|---|---|---|---|---|---|---|---|
| WT CNR2 | 16 | 0.3 (0.03) | 0.12 (0.02) | 5.0 | 5.1 (0.48) | 22.02 (4.00) | 39.75 (12.96) |
| WT A2aR | 16 | 0.3 (0.03) | 0.12 (0.02) | 4.0 | 5.8 (0.19) | 5.12 (0.59) | 5.52 (1.68) |
| TM6* CNR2 | 25 | 0.2 (0.02) | 0.32 (0.02) | 9.0 | 23.3 (1.00) | 17.02 (2.20) | 30.81 (7.71) |
| TM6* A2aR | 25 | 0.2 (0.02) | 0.32 (0.02) | 10 | 29.4 (0.98) | 5.16 (0.61) | 6.26 (1.86) |
The human GPCRs, A2aR and CNR2, were produced in the wild-type S. cerevisiae strain, BY4741 (WT), and the respiratory strain, TM6*. Cells were harvested at the time points indicated for preparation of membranes for functional analysis, as described in the Methods section. The mean growth rate (h-1) is calculated for the exponential growth phase (n = 2). The mean dry cell weight yield is reported with respect to the glucose consumed (n = 2). Harvest wet cell weight is reported in g, and the mean total membrane protein yield from these cells is reported (n = 3). Bmax and Kd values were calculated from the curves shown in Figure 5 (n = 3). In all cases the standard error of the mean is given in parentheses.