Localized contact formation by erythrocyte membranes: electrostatic effects

The topology of the contact seam of human erythrocytes adhered by dextran, an uncharged polymer, has been examined. Particular attention has been paid to the influence of electrostatic intermembrane interactions since their magnitude and range can be accurately estimated. Normal cells formed a continuous seam, whereas erythrocytes with pronase-modified glycocalices formed localized contact points on adhesion in 72 kDa dextran in buffered 145 mM NaCl. The dependence of the inter-contact distance lambda on dextran concentration [D] over the range 2–6% w/v, was given by lambda = C[D]-0.62, where C was a constant. The index of [D] was independent of dextran molecular mass over the range 20 to 450 kDa. The inter-contact distance for pronase-pretreated cells in 6% w/v 72 kDa dextran increased from 0.78 to 1.4 microns as [NaCl] was reduced through the range 145 to 90 mM and the suspending phase was maintained at isotonicity by using sorbitol to replace NaCl. The formation and lateral separation of the contact points are discussed from the perspective of linear interfacial instability theory. The theory allows a quantitative explanation for the experimentally observed dependence of inter-contact distance and of disturbance growth rate on change in electrostatic interaction. The results suggest that the dominant wavelength, determining the inter-contact distance, is established on approaching membranes when the layers of cell surface charge are separated by a perpendicular distance of < 14 nm (bilayer separation of 24 nm)