PROTEIN-KINASE-C SUBSTRATES AND GANGLIOSIDE INHIBITORS IN BOVINE MAMMARY NUCLEI

In cow mammary gland, unlike in other tissues, gangliosides (putative physiologic regulators of protein kinase C) may be distributed in nuclei and on the cell surface. This study was designed to determine whether gangliosides and the protein kinase C system (the enzyme and its substrate proteins) are present in cow mammary gland nuclei and to examine the effect of gangliosides detected in nuclei on protein phosphorylation catalyzed by protein kinase C. Gangliosides GM3, GD3, and GT1b were detected in the highly purified nuclear fraction. The nuclear ganglioside pattern was different from those of whole tissue and cytosol, thereby suggesting the presence of the gangliosides in nuclei. Protein kinase C and its substrate proteins (120, 97, 56, 43, 38, and 36 kDa) were extracted by Triton X-100 treatment of nuclei. Both protein kinase C activity (histone phosphorylation) and the nuclear substrate phosphorylation were effectively inhibited by the three gangliosides. Of the gangliosides, GT1b was the most potent in inhibiting phosphorylation, followed by GD3 and GM3. These results suggest that signal transduction mediated by protein kinase C in cow mammary gland nuclei may be regulated by gangliosides.