Fig. 6. Enzymatic properties of MsGC-ß3 compared to MsGC-ß3
C338.
COS-7 cells were transiently transfected with the plasmids indicated and
assayed for guanylyl cyclase activity with GTP concentrations of 0.1 mmol
l-1 to 5 mmol l-1 in the presence of 4 mmol
l-1 MgCl2 (A,B) or 4 mmol l-1
MnCl2 (C,D). For each concentration the reaction was allowed to
proceed for 5, 15 and 30 min and linear regression of these values used to
calculate a rate of reaction. The values were also plotted as double
reciprocal plots (B,D), which demonstrate that both MsGC-ß3 and
MsGC-ß3
C338 yield Hill coefficients of 1.0. Values for the
Vmax and Km were calculated from the
untransformed data using the following equation:
Rate=(Vmaxx[GTP])/(Km+[GTP])
using GraphPad Prism 3.0 and are shown in
Table 3.