Fluorescence polarization order parameters and phase transitions in lipids and lipoproteins

Fluorescence polarization measurements with 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to monitor phase changes in sonicated dispersions of triacylglycerols, cholesteryl esters and phospholipids. Lipid transitions to a fluid state were detected in a novel way by noting the temperature, t0.08, at which rs, the steady state anisotropy, was equal to 0.08. According to published equations (Van Blitterswijk , W.J., Van Hoeven , R.P. and Van der Meer , B.W. (1981) Biochim. Biophys. Acta 644, 323-332), this value for rs corresponds to a value of zero for S, a structural order parameter. Saturated and trans monounsaturated fatty triacylglycerols and distearoylphosphatidylcholine yielded t0.08 values in close agreement with transition temperatures found by differential scanning calorimetry (DSC), whereas cis unsaturated triacylglycerols displayed residual anisotropy, rs greater than 0.08, at temperatures above the DSC transition. The bent configuration of the cis double-bonded fatty acyl chains probably limits freedom of movement even in the liquid state when three such chains are bound to the glycerol molecule. Cholesteryl esters of 14:0, 18:0 and cis 18:1 fatty acids all showed rs greater than 0.08 above the DSC melting point. The difference in rotational freedom of DPH in triacylglycerol and cholesteryl esters even in the 'liquid' state explains the low t0.08 in the more fluid plasma VLDL and the contrastingly high t0.08 in plasma LDL, and HDL, which contain more cholesteryl ester an less triacylglycerol.