Dynamic properties of the haptenic site of lipid haptens in phosphatidylcholine membranes. Their relation to the phase transition of the host lattice.
The relation between the dynamic properties of the haptenic site of lipid haptens and the phase transition of the host lattice was investigated using head group spin-labeled phosphatidylethanolamines, that is, spin-label lipid haptens (Brûlet, P., and H. M. McConnell, 1976, Proc. Natl. Acad. Sci. USA., 73:2977-2981; Brûlet, P., and H. M. McConnell, 1977, Biochemistry, 16:1209-1217). The electron spin resonance (ESR) spectra of the lipid haptens in liposomal membranes showed three narrow resonance lines, whose widths and hyperfine splitting values suggested that the haptenic site, i.e., the spin-label moiety, should be exposed in the water phase. The line width of each peak depended on the host lipid species and on the incubation temperature. A temperature study using dipalmitoylphosphatidylcholine (DPPC) liposomes showed that the dynamic properties of the haptenic site were related to the main phase transition and the subphase transition of the host lattice but not to the prephase transition. The angular amplitudes of the tumbling motion of the haptenic site were estimated using oriented multibilayer systems. The angular amplitude of dipalmitoyl-phosphatidyl-N-[[N-(1-oxyl-2,2,6, 6-tetramethyl-4-piperidinyl)-carbamoyl]-methyl]-ethanolamine in DPPC membranes was 63 degrees at 2 degrees C, and it increased slightly with an increase in temperature regardless of the phase transition of the host lattice. The value for egg phosphatidylcholine (PC) at 25 degrees C was the same as for DPPC above its main phase transition temperature. Rotational correlation time analysis showed that the axial rotation of the haptenic site was preferable to the tumbling motion of the rotational axis, and the predominance depended on the phase transition, Lc----L beta' and P beta'----L alpha. Elongation of the spacer arm between the haptenic site and phosphate increased the angular amplitude of the tumbling motion but reduced the effect of the host lattice. Spin-label lipid haptens with unsaturated fatty acyl chains were distributed heterogeneously in DPPC membranes, whereas those with the same fatty acyl chain as the host lattice were distributed randomly. The ESR spectrum of a lipid hapten under its prephase transition temperature showed two components, broad and narrow. This suggests that at least two different domains, a hapten-rich domain and a hapten-poor one, may coexist in membranes. ESR measurements at various temperatures suggested that the haptenic site fraction in the hapten-rich domain decreased in part during the phase transition from L beta' to P beta', and disappeared completely in the La phase. The spatial mobility and lateral diffusion of lipid haptens will be discussed in greater detail.
Takeshita
K
K
Department of Health Chemistry, School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Utsumi
H
H
Hamada
A
A
D015060:1,2-Dipalmitoylphosphatidylcholine; D002462:Cell Membrane / Q000276:immunology; D002784:Cholesterol; D004578:Electron Spin Resonance Spectroscopy; D006241:Haptens; D008051:Lipid Bilayers; D008954:Models, Biological; D008968:Molecular Conformation; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D013113:Spin Labels
D006241:Haptens; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D013113:Spin Labels; D015060:1,2-Dipalmitoylphosphatidylcholine; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine; D002784:Cholesterol