Preparation and characterization of hormone sensitive, resealed erythrocyte ghosts

A method for preparing resealed turkey erythrocyte ghosts is described which utilizes hypotonic lysis and resealing following restoration of isotonicity. The resealed ghosts are isolated above 55% sucrose. The resealed ghosts are shown to be capable of maintaining high intracellular K⁺ concentrations in the presence of a low K⁺ extracellular environment. When ATP and an ATP regenerating system are included during the resealing stage, (R) (-) epinephrine and NaF stimulated cyclic AMP accumulation, which is linear for 20 min, can be demonstrated. The concentration of (R) (-) epinephrine producing a half maximal response in resealed ghosts is 1.0 ± 0.4 x 10^-6M. This is the same as that for (R) (-) epinephrine in the intact erythrocyte. The resealed ghosts are impermeable to Ca²⁺, but Ca²⁺ inhibition of cyclic AMP accumulation is noted if the divalent cation ionophore, A 23187, is present or if Ca²⁺ is included during the resealing stage.