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Journal of Lipid Research, Vol 31, 17-26, Copyright © 1990 by Lipid Research, Inc.
PA Kern, RA Martin, J Carty, IJ Goldberg and JM Ong
Postheparin plasma is a convenient source for the measurement of
lipoprotein lipase (LPL) in humans. Previous studies have focused on the
measurement of LPL catalytic activity, and have been unable to conveniently
measure the LPL protein or identify possibly different plasma forms of the
enzyme. Pre- and postheparin plasma was treated with a highly specific
antibody raised against bovine milk LPL and the immunoprecipitate was
analyzed by Western blotting. In normal subjects there were several species
of LPL in plasma. A 56 kD protein increased after heparin injection, and
likely represented active LPL. The anti- LPL antibody reacted specifically
with this 56 kD protein, and also reacted specifically with proteins at 52
kD, 69 kD, as well as a 20 kD breakdown product. In addition, using peptide
mapping, the 56 kD protein was structurally similar to the 52 and 69 kD LPL
proteins. The antibodies were affinity purified, biotinylated, and used to
quantitate LPL immunoreactive mass using an enzyme-linked immunosorbent
assay (ELISA). LPL immunoreactive mass was present in all subjects in
preheparin plasma. In postheparin plasma, five patients with type I
hyperlipoproteinemia displayed decreased LPL immunoreactive mass when
compared to normal subjects, although there was a wide range of specific
activity of the small amount of enzyme present. When the LPL from the
plasma of the patients was immunoprecipitated and Western blotted, there
was considerable heterogeneity in the appearance of the LPL forms, and an
overall decrease in LPL protein. Thus, several different immunoreactive LPL
proteins were present in pre- and postheparin plasma. In preheparin plasma,
as well as in patients with type I hyperlipoproteinemia, there was
decreased immunoreactive LPL protein, and the LPL protein that was present
was of low specific activity.
ARTICLES
Identification of lipoprotein lipase immunoreactive protein in pre- and postheparin plasma from normal subjects and patients with type I hyperlipoproteinemia
Department of Medicine, Cedars-Sinai Medical Center/UCLA 90048.
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