Calcium stimulation of procoagulant activity in human erythrocytes. ATP dependence and the effects of modifiers of stimulation and recovery.

The human erythrocyte membrane is generally considered to have no procoagulant activity. The normal membrane is characterized as having an asymmetric distribution of phospholipid species such that negatively charged and aminophospholipids are predominantly located on the inner leaflet of the membrane bilayer. Elevation of cytoplasmic Ca2+ in erythrocytes produces an assortment of biochemical and structural responses that include diminished phospholipid asymmetry and an elevation in procoagulant activity. Maintenance of the normal asymmetric distribution of phospholipid species is believed to be largely mediated by a phospholipid translocase mechanism. We have utilized a recently developed single-step kinetic assay of procoagulant activity to investigate the mechanisms of Ca2+ stimulation of procoagulant activity and recovery from the procoagulant state upon removal of Ca2+. This study demonstrated that stimulation of procoagulant activity by elevated cytoplasmic Ca2+ is greatly diminished in ATP-depleted erythrocytes. Phospholipid translocase inhibitors failed to fully inhibit recovery from the procoagulant state after removal of Ca2+. The data indicate that recovery of endogenous lipid from a procoagulant cofiguration may not be entirely mediated by the phospholipid translocase. Additionally, the data are inconsistent with the phospholipid translocase mediating the Ca(2+)-induced elevation of procoagulant activity, although the involvement of other protein(s) is indicated.