LATERAL DOMAIN FORMATION IN CHOLESTEROL PHOSPHOLIPID MONOLAYERS AS AFFECTED BY THE STEROL SIDE-CHAIN CONFORMATION

The interaction of side-chain variable cholesterol analogues with dipa lmitoylphosphatidylcholine (DPPC) or N-palmitoylsphingomyelin (N-PSPM) has been examined in monolayer membranes at the air/water interface. The sterols had either unbranched (n-series) or single methyl-branched (iso-series) side chains, with the length varying between 3 and 10 ca rbons (C3-C10). The efficacy of interaction between the sterols and th e phospholipids was evaluated based on the ability of the sterols to f orm condensed sterol/phospholipid domains in the phospholipid monolaye rs. Domain formation was detected with monolayer fluorescence microsco py using NBD-cholesterol as the fluorescent probe. In general, a side chain length of at least 5 carbons was necessary for the unbranched st erols to form visible sterol/phospholipid domains in DPPC or N-PSPM mi xed monolayers. With the iso-analogues, a side chain of at least 6 car bons was needed for sterol/phospholipid domains to form. The macroscop ic domains were stable up to a certain surface pressure (ranging from 1 to 12 mN/m). At this onset phase transformation pressure, the domain line boundary dissipated, and the monolayer entered into an apparent one phase state (no clearly visible lateral domains). However, with so me DPPC monolayers containing short chain sterols (n-C3, n-C4, n-C5, a nd i-C5), a new condensed phase appeared to form (at 20 mol%) when the monolayer was compressed beyond the phase transformation pressure. Th ese precipitates formed at surface pressures between 6-8.3 mN/m, were clearly observable up to at least 30 mN/m. When the monolayers contain ing these four sterols were allowed to expand, the condensed precipita tes dissolved at the same pressure at which they were formed during mo nolayer compression. No condensed precipitates were observed with thes e sterols in corresponding N-PSPM monolayers. Taken together, the resu lts of this study emphasize the importance of the length and conformat ion of the cholesterol side chain in determining the efficacy of stero l/phospholipid interaction in model membranes. The major difference be tween DPPC and N-PSPM monolayers at different sterol compositions was mainly the lateral distribution and the size of the domains as well as the onset phase transformation pressure intervals.