Lipid Structure and Membranes, Summary i Norman Albon July 2009

Contents:
Lipids,Membranes,Cells,Crystals,X-Ray Diffraction,Thermal Analysis, Raman
Spectra,Crystallography,Molecular Structures,Lecithin,Cardiolipin,Molecular Motion.

Phospholipids are found in cell membranes of complex life forms and disperse in water giving
bimolecular structures that isolate water regions. Research on lipid structures at a molecular and atomic
level is described. Among ‘widely accepted models’ for membranes (Wikipedia) is the fluid mosaic
model of 1972, that regards membranes to be ‘two dimensional liquids’ where all lipid and protein
molecules diffuse more or less freely in a space of 10 nm, being held together by non covalent
interactions between the chains.
Kinetic theory of liquids regards differences between solids and liquids as in their degree of order. In
1943, FRENKEL wrote ‘an understanding of the principles of the kinetic theory of liquids seems to
have reached thus far to a somewhat narrow circle of physicists and chemists, whereas it is of vital
importance to all those who have to deal with matters in the condensed state’

Research on lipids and their structural transitions, topics not in the fluid mosaic theory, was described
in the Journal of Chemical Physics (1983). When lecithin is added to water, as in a membrane, the
chains form a hexagonal structure with chain axes 4.85Å (0.48nm) apart. Sometimes described as ‘self
assembly’, it is like crystal growth. On heating, the chains become disordered at a transition that is very
sharp for pure compounds. The area per molecule (of two chains) is 48Ų below this transition and
~60Ų above. Crystallography of triglycerides had shown that Carboxy-ester (CE) groups, formed from
Carbonyl (C=O) and Glycerol, are planar and rigid, whilst Carbon-Carbon and ester bonds such as P-
O-C permit rotation. The hydrogen atoms are most easily excited on heating. Lipid molecules fold with
parallel chains attached to CE groups. In 1983, the greater stability of hexagonal chain packing in
lipids over that in single hydrocarbons was attributed to the CE-C-CE group. Disordered chains remain
anchored to that group. To the statement that the lipid membrane is held together by non covalent
forces between the chains, we add that the chains in each molecule are also held together by the much
stronger covalent bonds of the CE-C-CE groups. The molecular layers are flexible rather than fluid.

In phospholipid molecules, Phosphate (PO4) joins polar groups and Diglyceride. 1,2 sn dipalmitoyl
glyceride (DPG) yields anhydrous crystals with chains in the O_|_ subcell for which extensive
diffraction data was measured at 20˚C and 87K. THERMAL ANALYSIS (1986) indicated that the
crystal, hexagonal and disordered chain phases of this compound are closely related in structure.
Hexagonal and disordered chain phases of diglyceride groups in phospholipids are unlikely to differ,
the transition between these phases is at 41.55˚C in lecithin water systems and at 54˚C for DPG
(anhydrous).
Orientation of the DPG molecule in the unit cell was established by SINGLE CRYSTAL RAMAN
SPECTRA at 20˚C in 1984. Strong features from C-H bonds and CH 2 groups confirmed the major role
of hydrogen-hydrogen contacts in molecular packing. Sharp spectra from relatively few hydrogen
atoms in the methyl groups indicate low thermal motion of the molecules at the measurement
temperature, as the chain disorder transition for the crystal at 69.6˚C is ~50˚C higher than that of the
crystals studied by Raman spectra.
The CRYSTAL STRUCTURE of DPG (1993) at 87K with R= 0.0256 gave precise details of the
molecular conformation and packing and confirmed the main conclusions from Raman spectra. The
structures at 20˚ and 87K are identical except for thermal vibrations. In crystals, DPG molecules are
oriented in rows along the unit cell diagonal of axes a-b, with the chains in the direction of axis c. The
asymmetric carbon atom that joins the CE groups and the molecules all have the same orientation.

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On heating DPG crystals from 87 to 293K, axis b expands by 0.294Å, 10 times that of axis a. This can
be attributed to an increase in the methylene twisting-rocking and wagging vibrations seen in Raman
spectra. Closely packed hydrogen atoms in the crystal have 60 contacts of 2.43-2.50Å and are attached
to regular carbon skeletons. These move apart at the transition to disordered chains when heated to
343K. Conformation angles now deviate from 180˚, due to rotation about carbon-carbon bonds.
Hydrogen bonds are absent from this phase and the hexagonal phase formed on cooling. On further
cooling to 20-30˚C, for DPG, not phospholipids, hexagonal transforms to the crystal phase with a
hydrogen bond, and shorter b axis. Motion of the hydrogen atoms results in a rounded chain profile
perpendicular to the chain axes. In hexagonal packing a slight degree of chain distortion gives the
diffraction observed at 4.2Å, mainly from carbon atoms, and a large number of hydrogen-hydrogen
contacts. The increased ease of formation and stability of hexagonal chain packing in diglycerides can
be readily demonstrated with pairs of linked cylinders.

The structure of Lecithin hydrate crystals does not provide a membrane model as the very stable double
layer of phosphorylcholine (PC) dominates although the chains still pack with parallel axes. The
molecular folding is similar to that of triglycerides.
MEMBRANE MODELS were obtained by replacing the chains in the DPG structure with hexagonal
chain packing, (so now axis b is longer by 0.84Å) and adding a Phosphate polar group. That polar
group will be held in the water layer by two flexible ester bonds and only exert weak forces on glycerol
carbon sn1. The increase in chain volume is small and easily taken up by C-C bond rotations with the
CE-C-CE group unaltered.
On heating purified lecithin to 314.55K, there is a sharp transition to disordered chains. The observed
volume change of 7-10% does not require a significant alteration of the link between the chains.

A variation in molecular tilt and the related area per molecule, occurs in the hexagonal phase. For
LECITHIN with a phosphorylcholine head group the area is independent of chain length for lipids from
C16 and C14 fatty acids. Models for these structures using the measured area have a slightly larger
chain tilt than that in DPG crystals. PC is a large group, small groups might involve zero chain tilt with
minimum area but we have no data for phosphatidyl glycerol, a small head group. A related compound,
CARDIOLIPIN or diphosphatidylglycerol has four chains joined by diphosphatidyl with covalent
bonds. These have known atomic parameters. The model proposed for lecithin but with zero chain tilt
fits cardiolipin precisely. Maximum levels of cardiolipin are found in biological materials that require
another, two chain, phospholipid molecule for a membrane.

The presence of vacancies in the structure from adjacent areas with different orientations, is proposed
as the mechanism of molecular movement in each single layer. That results in sites of dimensions
similar to that of the fluid mosaic model. This would not accommodate layers composed entirely of
cardiolipin, but is valid if another smaller phospholipid molecule is present.

Experimental work has been published in scientific journals except for the mechanism for molecular
motion in layers of double hexagonal cylinders, a simple development of solid state theory.

Spreading of hydrocarbons on a water surface and coalescence as droplets in water can be commonly
observed phenomena. How a simple molecular structure ensures the formation of such very thin stable
hydrocarbon layers in water, essential for living cells is remarkable. In a fuller account, simple
explanations of the structures and of the methods used in the research are given to add to more
technical research papers.
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