Receptors for the phorbol ester tumour promoters.

The phorbol esters are potent tumour promotors in mouse skin and have profound effects on a wide variety of biological systems. Using the derivative [20-3H]phorbol 12,13-dibutyrate ([3H]PDBu), we demonstrated that specific receptors for the phorbol esters exist which mediate many of these biological effects. The receptors represent a complex between phospholipids and protein kinase C, an enzyme first identified by Nishizuka and coworkers. Considerable evidence indicates heterogeneity in the pharmacology of the biological responses to the phorbol esters. Likewise, multiple subclasses of binding sites have been observed. Differences in the phospholipids associated with protein kinase C may account for this heterogeneity, affecting both absolute and relative affinities. An endogenous analogue of the phorbol esters had been predicted from the high evolutionary conservation of the receptor. Nishizuka and coworkers have reported that diacylglycerols appear to be natural activators of protein kinase C. We find that diacylglyerols competitively inhibit phorbol ester binding, consistent with their acting at the same site on the enzyme. Likewise, by binding analysis, we can demonstrate a 1:1 stoichiometry between D-1-2-diacylglycerol and the receptor. The potencies of diacylglycerols for binding reflect their local concentration in the phospholipids. The K1 for diolein is approximately 0.1%. Relative to the corresponding phorbol esters, the potencies of the diacylglycerols are somewhat (17-fold) to markedly (30 000-fold) lower. We conclude that factors affecting phospholipid-receptor interactions or diacylglycerol production are of potential importance in the promotion process.