Influence of red cell surface charge on red cell membrane curvature.

The wellknown biconcave shape of resting RBC reflects an intrinsic overall negative spontaneous membrane curvature. It depends to a high degree on the integrity of the spectrin-actin-ankyrin-Band-3 hetero-complex. Alterations of this complex have previously been shown to be associated with shape transitions, which have been abolished by treatment with enzymes reducing the surface charge of the RBC. In this report we show that treatment of erythrocytes with enzymes reducing the surface charge (e.g. neuraminidase, trypsin, chymotrypsin and pronase), consistently exerts a "stomatocytogenic" effect, i.e. it reduces mean mean curvature. Also the time dependency for the charge reduction and for the correlated decrease of mean mean curvature is shown. So-called stomatocytogenic agents (e.g. clorpromazine, tetracain and triton X100) and so-called echinocytogenic agents (e.g. dinitrophenol and Na-salicylate) are known to change membrane curvature in a dose dependent manner. It is further shown that by prior reduction of surface charge by various enzymes the dose response curves of all stomatocytogenic and echinocytogenic agents tested is shifted towards higher degrees of stomatocytosis or lesser degrees of echinocytosis. The data show, that in RBC pronounced curvature influences are produced by the surface charge located on the sialic acid residues in the glycocalix.