Inhibition by sulfatide of 21-kDa protein phosphorylation by protein kinase C in cow mammary gland and its reversal by phosphatidylserine.
The effect of sulfatide, a sulfated sphingolipid, on phosphorylation of endogenous proteins by protein kinase C (PKC) was examined in cow mammary gland. Several proteins, including 21-kDa, 43-kDa and 56-kDa proteins in the cytosolic fraction, were found to be substrates for PKC by phosphorylation in the absence or presence of the cofactors 1-oleoyl-2-acetyl-sn-glycerol (OAG), phosphatidylserine (PS) and Ca2+. Sulfatide inhibited the 21-kDa phosphorylation, whereas it enhanced the 56-kDa and 43-kDa phosphorylation. Experiments were then conducted to examine whether other sphingolipids, including sphingosine, dihydrosphingosine, ceramides, galactocerebrosides, psychosine and sphingomyelin, modulated phosphorylation of the PKC substrates. Sphingosine, dihydrosphingosine and psychosine did not inhibit the 21-kDa phosphorylation; however, they enhanced the 56-kDa and 43-kDa phosphorylation. Ceramides, galactocerebrosides and sphingomyelin did not inhibit the 21-kDa or enhance the 56-kDa and 43-kDa phosphorylation. The inhibition by sulfatide of the 21-kDa phosphorylation was reversed by excess addition of PS, but not by OAG or Ca2+; whereas the enhancement by sulfatide, as well as sphingosine, dihydrosphingosine and psychosine, of 56-kDa and 43-kDa phosphorylation was not affected by PS, OAG or Ca2+. It is suggested that sulfatide is involved in the regulation of PKC-dependent phosphorylation by modulating the association of PKC substrates, in particular the 21-kDa protein, with membrane phospholipids in cow mammary gland.
National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
Norio
N
Katoh
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D004075:Diglycerides / Q000378:metabolism; D005260:Female; D008321:Mammary Glands, Animal / Q000187:drug effects* / Q000201:enzymology; D008970:Molecular Weight; D018994:Myosin Light Chains / Q000378:metabolism*; D010718:Phosphatidylserines / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D013107:Sphingolipids / Q000378:metabolism; D013433:Sulfoglycosphingolipids / Q000494:pharmacology*
D004075:Diglycerides; D018994:Myosin Light Chains; D010718:Phosphatidylserines; D013107:Sphingolipids; D013433:Sulfoglycosphingolipids; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D002118:Calcium