Interdigitated gel phase bilayers formed by unsaturated synthetic and bacterial glycerolipids in the presence of polymyxin B and glycerol

Abstract

The ability of synthetic phosphoglycerolipids with a cis mono-unsaturated acyl chain in the 2-position and a saturated chain in the 1-position of glycerol to form interdigitated gel phase bilayers in the presence of amphipathic substances was monitored using a fatty acid spin label, 16-doxylstearic acid, and phosphatidylglycerol spin label containing 16-doxylstcaric acid. These spin labels become significantly more motionally restricted in an interdigitated gel phase bilayer than in a non-interdigitated gel phase bilayer. The results indicated that polymyxin B and polymyxin B nonapeptide caused interdigitation of 1-palmitoyl,2-oleoyl-phosphatidylglycerol (POPG) and glycerol caused interdigitation of 1-stearoyl,2-olcoyl-phosphatidylcholine (SOPC), similar to their effects on disaturated lipids. The fluidity gradient present in non-interdigitated gel phase bilayers was abolished. However, glycerol did not cause POPG to become interdigitated, in contrast to SOPC. We reported earlier that there is a kinetic barrier to interdigitation of saturated PG in the presence of glycerol, in contrast to saturated PC. This barrier is even greater for the unsaturated species of PG. Furthermore, these compounds lowered the gel to liquid-crystalline phase transition temperatures of the unsaturated lipids more than of saturated lipids suggesting that the interdigitated bilayer of the former may be less ordered or less stable than that of the latter. Since polymyxin B is an antibiotic we also examined its effect on a lipid extract from the Gram-negative bacteria Pseudomonas aeruginosa in order to assess whether interdigitation might be involved in its mechanism of bactericidal or bacteriostatic effect. Polymyxin B and polymyxin B nonapeptide also caused motional restriction of a small percentage (about 13% at −2°C and 25% at −14°C for polymyxin B) of the spin label in the lipid extract at low temperatures, where the lipid is in the gel phase, consistent with formation of a small domain of interdigitated bilayer lipid. However, the degree of immobilization was less than that in the interdigitated bilayers of the synthetic unsaturated lipids. This may be a result of the heterogeneous nature of the lipids in the extract. However, it cannot be ruled out that the motional restriction of the spin label in this extract may be caused by something other than interdigitation. Thus the results with the lipid extract are less conclusive of interdigitation than for the synthetic lipids. A motionally restricted population was not detectable at higher temperatures. This study shows that a cis-double bond in the acyl chain of phosphoglycerolipids does not prevent formation of an interdigitated gel phase bilayer. However, such lipids are in the liquid-crystalline phase at physiological temperature and thus will not form interdigitated domains in biological membranes.