Chemo-mechanical leak formation in human erythrocytes upon exposure to a water-soluble carbodiimide followed by very mild shear stress. I. Basic characteristics of the process

Human erythrocytes treated with low concentrations (1-5 mM) of the carboxyl group-modifying reagent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) lose their native deformability in parallel with extensive cross-linking of the membrane skeleton. After treatment with higher (5-40 mM) concentrations of the reagent the cells develop a hitherto undescribed property: when subjected to even very low shear stresses (resuspension after packing by centrifugation or viscometric shearing at up to 4 s-1) they become highly leaky to ions, lose their K+ with a half-time of about 5 min and subsequently undergo hemolysis. Lysis is not accompanied by cell fragmentation as occurs with mechanical hemolysis, but is colloid-osmotic, due to the formation of aqueous membrane leaks with an apparent radius of about 3 nm. Leakiness and lysis affect an increasing fraction of the cell population, in relation to (a) the concentration of EDC applied, (b) the shearing intensity, and (c) particularly, the hematocrit during shearing. The physical parameter determining the mechanical component of this 'chemo-mechanical' leak formation is not predominantly the shear stress. Rather, cell-cell interactions of as yet undefined nature seem to be involved. The analysis of chemo-mechanical leak formation may provide interesting insights into the influence of mechanical forces on membranes.