Previous in vitro experiments have shown that the phorbol-like diterpenes 12-deoxyphorbol 13-isobutyrate (dPB), and possibly mezerein, have multiple biological target sites which differ from one another in apparent affinity for dPB by 12.5-780 fold and for mezerein by 24-fold. These two compounds are thus very important ligands because of their potential PKC isotype-selectivity. In the present study they were found to have binding affinities differing by a maximum of only 1.6-fold among recombinant protein kinase C (PKC) isotypes alpha, beta 1, beta 2, and gamma (the "A-group") in a [3H]phorbol dibutyrate binding assay. The apparent Ki's were 92-140 nM for dPB and 68-92 nM for mezerein. Our results are consistent with short-term 12-deoxyphorbol ester-induced mouse skin inflammation being mediated at least in part by one or more A-group PKC isotypes. The data also indicate that the pharmacologically distinguishable target sites previously established for mezerein and dPB must include one or more binding sites not found in the A-group of PKC isotypes and that mezerein has a high-affinity, non-A-group target site in brain.