Separation from protein kinase C — a calcium‐independent TPA‐activated phosphorylating system

Abstract
A calcium-independent but 12-O-tetradecanoylphorbol-13-acetate (TPA)- or diacylglycerol-activated phospholipid-dependent phosphorylating activity has been separated from protein kinase C. This has been made possible by employing calcium-dependent hydrophobic interaction chromatography. The material bound to phenyl-Sepharose in the presence of calcium at low ionic strength was eluted with EGTA and was protein kinase C. While the unbound material passing through the phenyl-Sepharose column showed no appreciable protein kinase C activity, instead it had a high phosphorylating activity manifested in the absence of calcium and in the presence of TPA plus phospholipid. The identification of this phosphorylating activity, distinct from protein kinase C, leads to important clues to cellular responses monitored by TPA in the absence of calcium.