ATP analogues induce membrane permeabilization in transformed mouse fibroblasts

The mechanism underlying ATP-induced permeabilization of transformed mouse fibroblasts was studied by using nonhydrolyzable analogues of ATP. Incubation of 3T6 cells with 0.6 mM of either ATP, 5′-adenylyl imidodiphosphate (p[NH]ppA) or adenosine 5′-[β,γ-methylene]triphosphate (p[CH₂]ppA) resulted in an increase of 17-, 8- or 5-times, respectively, in the cell membrane permeability, measured by the efflux of normally impermeant metabolites from the cells. The induced cell permeabilization was preceded by a reduction in the membrane potential (Δψ), determined according to the distribution of the cation tetraphenylphosphonium (TPP⁺) between the cells and the medium. Reduction of 26, 18 and 13 mV in Δψ was exerted by 0.6 mM of either ATP, p[NH]ppA or p[CH₂]ppA, respectively. In 3T3 cells the untransformed counterparts of 3T6 cells, neither reduction of Δψ, nor alterations in membrane permeability were exerted by either ATP or by its analogues. The data indicate that the dissociation of the β,γ-phosphate bond is not essential for membrane permeabilization by external ATP, implying that the binding of ATP to the cell surface of transformed cells is sufficient to initiate the permeabilization process. The data also suggest that Δψ is involved in the control of membrane permeability.