Vesicles isolated from ATP-depleted erythrocytes and out of thrombocyte-rich plasma.

When human erythrocytes are depleted of endogenous ATP they release spectrin-free vesicles as a light vesicle fraction [Lutz et al: J Cell Biol 73: 548, 1977] and chains of rounded vesicles as well as flattened myelin forms in a heavy vesicle fraction. The heavy fraction retains some spectrin, and glycophorin is partially degraded. The release of both types of fragments is not dependent upon added Ca+2, and 50 micrometer EGTA does not prevent the vesicle release. Concomitant with vesicle release, a large fraction of the major protein components of the cell is found in disulfide-bonded aggregates. A protocol is outlined to recover erythrocyte-specific fragments from thrombocyte-rich plasma. It allows detection of spectrin-free vesicles in whole blood stored under blood bank conditions for 12 days. In freshly drawn blood no such vesicles are observed, but particles are obtained that are different from thrombocyte fragments and that show a prominent glycoprotein running slightly faster than glycophorin.