We have previously found the 78-kDa glucose-regulated protein (Grp78) to be a subunit of protein phosphatase1(PP1)gamma2. To determine the role of Grp78 in PP1gamma2 holoenzyme, we compared the two forms of this enzyme, PP1gamma2 holoenzyme containing Grp78 and Grp78-dissociated PP1gamma2 in rat testes in terms of their kinetic constants and sensitivities to inhibitors of this enzyme. The enzymatic activity of the Grp78-dissociated enzyme was much lower at whole range of concentrations of a substrate (phosphorylase a) than that of the holoenzyme; the Km value was about ten-fold higher in Grp78-dissociated enzyme than in holoenzyme, while the Vmax was similar. IC50s of the Grp78-dissociated enzyme for three inhibitors (microcystin-LR, inhibitor-2, and okadaic acid) were more than ten-fold higher than those of the holoenzyme. These results indicate that the Grp78 subunit modulates the activity of PP1gamma2 through its actions to control the binding of substrates or inhibitors to PP1gamma2.
Copyright 1999 Academic Press.