The identification and purification of a mammalian-like protein kinase C in the yeast Saccharomyces cerevisiae.
We have purified a yeast protein kinase that is phospholipid-dependent and activated by Diacylglycerol (DAG) in the presence of Ca2+ or by the tumour-promoting agent tetradecanoyl-phorbol acetate (TPA). The properties of this enzyme are similar to those of the mammalian protein kinase C (PKC). The enzyme was purified using chromatography on DEAE-cellulose followed by hydroxylapatite. The latter chromatography separated the activity to three distinguishable sub-species, analogous to the mammalian PKC isoenzymes. The fractions enriched in PKC activity contain proteins that specifically bind TPA, are specifically phosphorylated in the presence of DAG and recognized by anti-mammalian PKC antibodies.
Department of Genetics, University of Leicester, U.K.
A J
AJ
Simon
Y
Y
Milner
S P
SP
Saville
A
A
Dvir
D
D
Mochly-Rosen
E
E
Orr
D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002845:Chromatography / Q000379:methods; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D004075:Diglycerides / Q000494:pharmacology; D017886:Durapatite; D004789:Enzyme Activation; D006882:Hydroxyapatites; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008322:Mammals; D008970:Molecular Weight; D010766:Phosphorylation; D011493:Protein Kinase C / Q000302:isolation & purification* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism / Q000494:pharmacology
D004075:Diglycerides; D006882:Hydroxyapatites; D007527:Isoenzymes; C038237:1-oleoyl-2-acetylglycerol; D017886:Durapatite; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate