Journal of Lipid Research, Vol 30, 765-771, Copyright © 1989 by Lipid Research, Inc.
Detection of vesicular lipoproteins in lecithin:cholesterol acyltransferase-deficient plasma by 1H-NMR spectroscopy
YI Parmar, DL Sparks, J Frohlich, PR Cullis and PH Pritchard
Department of Pathology, University of British Columbia, Vancouver, Canada.
The proton NMR spectra of the N-methyl choline region of normal and
lecithin:cholesterol acyltransferase (LCAT)-deficient lipoproteins and of
egg yolk phosphatidylcholine-cholesterol 55:45 (mol %) vesicle mixtures
have been examined in the presence and absence of manganous sulfate as a
line-broadening reagent. Manganous ions quenched all of the signal arising
from normal lipoproteins and only part of the vesicle signal corresponding
to the outer monolayer. There was no net loss of vesicular phospholipid
when vesicles were added to normal lipoproteins and as little as 5% (or 100
micrograms) of the vesicular phospholipid could be detected and quantitated
in the mixture of lipoproteins. Similar experiments performed on plasma
lipoproteins from an LCAT-deficient patient indicated that 42% of the
phospholipid was associated with vesicular lipoproteins. These experiments
demonstrate that this technique can be used to detect and quantify small
amounts of vesicular structures directly in a mixture of micellar
lipoproteins.
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