Influence of the length of the spacer on the partitioning properties of amphiphilic fluorescent membrane probes.

Four fluorescent diphenylhexatriene derivatives were considered as membrane probes, namely two ammonium compounds, 3-(diphenylhexatrienyl)propyltrimethylammonium (TMAP-DPH) and 22-(diphenylhexatrienyl)docosyltrimethylammonium (LcTMA-DPH), and two phospholipids, 1-palmitoyl-2-[3-(diphenylhexatrienyl)propanoyl]-sn-glyc ero-3-phosphocholine (DPHpPC) and 1-palmitoyl-2-[21-(diphenylhexatrienyl)henicosanoyl]-sn-phos phocholine (LcDPHpPC). For each pair, the molecules differ by the length of the polymethylenic spacer between the fluorescent moiety and the polar head, so one pair comprises two short chain molecules (C3 spacer) and the other two long chain molecules (C21 or C22 spacer). The partitioning of these probes between gel and liquid crystalline phases of multilamellar vesicles with binary composition (DEPC and DSPC) was measured by a method based on fluorescence anisotropy. The partitioning was shown to depend strongly on the length of the spacer. Short chain probes preferably partition into fluid phases (Kf/s = 1.7 +/- 0.3 for TMAP-DPH; 2.6 +/- 0.11 for DPHpPC), whereas long chain probes show a strong preferential partitioning for gel phases of the vesicles (Kf/s = 0.12 +/- 0.06 for LcTMA-DPH; 0.22 +/- 0.11 for LcDPHpPC). This strong partitioning may be explained by the interdigitation of the long polymethylenic chains across the mid-point of the lipid bilayer (I.E. Mehlhorn et al. (1988) Biochim. Biophys. Acta 939, 151-159), which is enhanced by the better packing provided by a gel phase.