Modulation by sphingosine of phosphorylation of substrate proteins by protein kinase C in nuclei from cow mammary gland.
Protein kinase C (PKC) is an enzyme activated by diacylglycerols such as 1-oleoyl-2-acetyl-sn-glycerol (OAG), phospholipids (in particular phosphatidylserine; PS) and Ca2+, which regulate a wide variety of intracellular functions by phosphorylating multiple substrate proteins and enzymes. The effect of sphingosine, the backbone moiety of sphingolipids, on PKC activity and phosphorylation of endogenous proteins catalyzed by PKC was investigated in nuclei of cow mammary gland. Sphingosine inhibited nuclear PKC activity when lysine-rich histone was used as the substrate. The sphingosine inhibition of the PKC activity was reversed by the excess addition of PS, but not by OAG or Ca2+. Several nuclear proteins, including 56-kDa, 43-kDa, 38-kDa and 36-kDa proteins, were shown to be substrates for PKC. Of the substrate proteins, the 38-kDa and 36-kDa proteins were identified as annexin I, the Ca2+/phospholipid-binding protein; the 56-kDa and 43-kDa proteins have not yet been identified. Sphingosine inhibited phosphorylation of the 56-kDa protein and the 36-kDa annexin I, whereas it enhanced that of the 43-kDa protein. The 38-kDa annexin I species was unaffected by sphingosine. As with the PKC activity, inhibition by sphingosine of phosphorylation of the 56-kDa protein and 36-kDa annexin I was reversed by the excess addition of PS, but not by OAG or Ca2+. In addition, by the excess addition of PS and not by OAG or Ca2+, the sphingosine-enhanced phosphorylation of the 43-kDa protein was reversed and returned to near the level in the absence of sphingosine. It is suggested that sphingosine is involved in the regulation of PKC-dependent phosphorylation in the nucleus by modulating the association of PKC or its substrates, particularly annexin I, with membrane phospholipids in cow mammary gland.
Katoh
Norio
N
National Institute of Animal Health, Ibaraki, Japan.
D000818:Animals; D017305:Annexin A1; D002118:Calcium / Q000378:metabolism; D002417:Cattle / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism; D005260:Female; D006657:Histones / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D013110:Sphingosine / Q000378:metabolism*
D017305:Annexin A1; D004075:Diglycerides; D006657:Histones; D009687:Nuclear Proteins; D010718:Phosphatidylserines; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D013110:Sphingosine; D002118:Calcium