Protein kinase C (PKC) from a human mcgakaryoblastic leukemic cell line (MEG-01) was resolved into two fractions by hydroxyapatitc column chromatography, which arc indistinguishable. from the brain type II (j?I//?II) and type III (u) subspecies, by biochemical and immunoblot analysis. In the presence of both phosphatidylserine and diacylglycerol, several free unsaturated fatty acids (FFA's), such as arachidonic, oleic, linoleic and linolenic acids, further enhanced the enzyme activation, and allowed the enzyme to exhibit almost full activity at nearly basal levels of Cat+ conccntration. The concentration of unsaturated FFA's giving rise to the maximum enzyme activation was around 2 x lO-s M. Palmitic and stearic acids were inactive. The result implies that, in addition to diacylglycerol, the receptor-mediated release of unsaturated FFA's from membrane phospholipids may also take part in the activation of PKC.