Tumor promoter 12-0-tetradecanoyl-phorbol-13-acetate (TPA) can reduce the Ca-transporting ability of Ca-ionophores in T lymphocytes: the involvement of intracellular heavy metal ions.

12-0-tetradecanoyl-phorbol-13-acetate (TPA) can significantly reduce the Ca-ionophore-induced rise in the intracellular calcium concentration (Cai) of T lymphocytes measured by quin2 or fura-2 fluorescence. This counteraction of TPA is maximal at a preincubation of 90 min at TPA concentrations higher than 20 nM. 45Ca uptake and efflux measurements directly indicate that TPA does not activate the calcium extrusion systems in thymocytes but impairs the Ca-transporting ability of Ca-ionophores. TPA causes no immobilization of the Ca-ionophores as it is demonstrated by the lack of significant changes in fluorescence and fluorescence polarisation of A23187 during TPA incubation. Similarly the energy transfer between the Tyr, Try groups of membrane proteins and A23187 shows no significant difference in control and TPA treated thymocytes. This indicates that A23187 is not in a membrane protein-bound form after TPA preincubation. The intracellular heavy metal chelator, N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) restores the ionophoretic ability of Ca-ionophores in TPA pretreated cells to the control level. Diacyl-glycerols also impair the Ca-transporting ability of Ca-ionophores. TPEN prevents this effect as well. These findings suggest that TPA and diacyl-glycerols may cause an increase in the availability of intracellular heavy metal ions. Our results may reflect a new, physiologically important mechanism of the action of diacyl-glycerols and phorbol esters.