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Biophys J, February 2000, p. 874-885, Vol. 78, No. 2

*The Research Institute, The Hospital for Sick Children,
Toronto M5G 1X8, Canada and
Department of
Laboratory Medicine and Pathobiology, University of Toronto,
Toronto, Ontario M5G 1L5, Canada
The two glycosphingolipids galactosylceramide (GalC) and
its sulfated form, cerebroside sulfate (CBS), are present at high concentrations in the multilayered myelin sheath and are involved in
carbohydrate-carbohydrate interactions between the lipid headgroups. In
order to study the structure of the complex of these two glycolipids by
Fourier transform infrared (FTIR) spectroscopy, GalC dispersions were
combined with CBS dispersions in the presence and absence of
Ca2+. The FTIR spectra indicated that a strong interaction
occurred between these glycolipids even in the absence of
Ca2+. The interaction resulted in dehydration of the
sulfate, changes in the intermolecular hydrogen bonding interactions of
the sugar and other oxygens, decreased intermolecular hydrogen bonding
of the amide C==O of GalC and dehydration of the amide region of one or both of the lipids in the mixture, and disordering of the
hydrocarbon chains of both lipids. The spectra also show that
Ca2+ interacts with the sulfate of CBS. Although they do
not reveal which other groups of CBS and GalC interact with
Ca2+ or which groups participate in the interaction between
the two lipids, they do show that the sulfate is not directly involved in interaction with GalC, since it can still bind to Ca2+
in the mixture. The interaction between these two lipids could be
either a lateral cis interaction in the same bilayer or
a trans interaction between apposed bilayers. The type
of interaction between the lipids, cis or
trans, was investigated using fluorescent and spin-label
probes and anti-glycolipid antibodies. The results confirmed a strong
interaction between the GalC and the CBS microstructures. They
suggested further that this interaction caused the CBS microstructures to be disrupted so that CBS formed a single bilayer around the GalC
multilayered microstructures, thus sequestering GalC from the external
aqueous phase. Thus the CBS and GalC interacted via a
trans interaction across apposed bilayers, which
resulted in dehydration of the headgroup and interface region of both
lipid bilayers. The strong interaction between these lipids may be
involved in stabilization of the myelin sheath.
Biophys J, February 2000, p. 874-885, Vol. 78, No. 2
© 2000 by the Biophysical Society 0006-3495/00/02/874/12 $2.00
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