Spectrin-phospholipid interaction. A monolayer study

Abstract

1.(1) The interaction of synthetic and natural phospholipids with spectrin, purified from human erythrocyte membranes, was studied using the monolayer technique at constant surface pressure. Spectrin penetration into the lipid monolayer was recorded as the rate of surface area increase on a two-compartment trough.

2. (2) High spectrin penetration rates were observed with negatively charged phospholipids while zwitterionic or neutral lipids showed only poor spectrin affinity. This penetration rate was strongly affected by the subphase pH. At pH 5.5, maximal pentration rates were observed for phosphatidylglycerol and phosphatidylserine but not for phosphatidylcholine.

3. (3) In comparing the penetration rates for phospholipids with a natural fatty acid composition and the dimyristoyl species of phosphatidic acid, phosphatidylglycerol, phosphatidylserine and phosphatidylcholine, the lipid fatty acid composition proved to be an important parameter. The differences are correlated with the area per lipid molecule.

4. (4) Other parameters affecting the area per lipid molecule such as surface pressure, pH and salt concentration also strongly influenced spectrin penetration rates for negatively charged phospholipids. Spectrin penetration into phosphatidylcholine monolayers is only slightly affected by variation of these conditions.

5. (5) The effect of Ca2+ on spectrin-lipid interactions was studied for several phosphatidylglycerol and phosphatidylserine species. Both lipids condensed upon the addition of Ca2+, but only in the case of the phosphatidylserine was this accompanied by extrusion of the spectrin from the interface, which is in agreement with earlier calorimetric experiments with bilayer systems of analogous composition (Mombers, C., Verkleij, A.J., de Gier, J. and van Deenen, L.L.M. (1979) Biochim. Biophys. Acta 551, 271–281). For this phenomenon a model is presented.