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J Biol Chem, Vol. 275, Issue 12, 9019-9025, March 24, 2000
From the Previous studies have provided detailed
information on the formation of spherical high density lipoproteins
(HDL) containing apolipoprotein (apo) A-I but no apoA-II (A-I HDL) by
an lecithin:cholesterol acyltransferase (LCAT)-mediated process. In
this study we have investigated the formation of spherical HDL
containing both apoA-I and apoA-II (A-I/A-II HDL). Incubations were
carried out containing discoidal A-I reconstituted HDL (rHDL),
discoidal A-II rHDL, and low density lipoproteins in the absence or
presence of LCAT. After the incubation, the rHDL were reisolated and
subjected to immunoaffinity chromatography to determine whether
A-I/A-II rHDL were formed. In the absence of LCAT, the majority of the
rHDL remained as either A-I rHDL or A-II rHDL, with only a small amount
of A-I/A-II rHDL present. By contrast, when LCAT was present, a
substantial proportion of the reisolated rHDL were A-I/A-II rHDL. The
identity of the particles was confirmed using apoA-I rocket
electrophoresis. The formation of the A-I/A-II rHDL was influenced by
the relative concentrations of the precursor discoidal A-I and A-II
rHDL. The A-I/A-II rHDL included several populations of HDL-sized
particles; the predominant population having a Stokes' diameter of 9.9 nm. The particles were spherical in shape and had an electrophoretic mobility slightly slower than that of the
Formation of Spherical, Reconstituted High Density Lipoproteins
Containing Both Apolipoproteins A-I and A-II Is Mediated by
Lecithin:Cholesterol Acyltransferase*
§¶,§,
, and§
The University of Adelaide, Department of
Medicine, Royal Adelaide Hospital, North Terrace, Adelaide,
South Australia 5000, Australia, § Lipid Research
Laboratory, Hanson Centre for Cancer Research, Institute of Medical and
Veterinary Science, Frome Road, Adelaide,
South Australia 5000, Australia, The Cardiovascular
Investigational Unit, Royal Adelaide Hospital, North Terrace, Adelaide,
South Australia 5000, Australia
-migrating HDL in human plasma. The apoA-I:apoA-II molar ratio of the A-I/A-II rHDL was 0.7:1.
Their major lipid constituents were phospholipids, unesterified cholesterol, and cholesteryl esters. The results presented are consistent with LCAT promoting fusion of the A-I rHDL and A-II rHDL to
form spherical A-I/A-II rHDL. We suggest that this process may be an
important source of A-I/A-II HDL in human plasma.
*
This work was supported by a Grant G 97A 4966 from the
National Heart Foundation of Australia.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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