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J. Biol. Chem., Vol. 262, Issue 19, 8944-8947, Jul, 1987
S Lamon-Fava, JM Ordovas, G Mandel, TM Forte, RH Goodman and EJ Schaefer
Apolipoprotein A-I (apoA-I) is the major protein constituent of plasma high
density lipoproteins (HDL). To examine apoA-I processing and secretion, the
human apoA-I gene (2.2-kilobase PstI-PstI fragment) linked to the mouse
metallothionein promoter was transfected by electroporation into NIH 3T3
fibroblasts along with the plasmid pSV2 neo, which confers neomycin
resistance. Transfected cells were selected for neomycin resistance and
screened for the ability to produce apoA-I by enzyme-linked immunosorbent
assay. In the absence of lipids in the medium, selected 3T3 cells secreted
apoA-I, mainly in the proprotein form, at density greater than 1.25 g/ml.
Following incubation of cells with lipids, and subsequent washing with
lipid-free medium, apoA-I was recovered in the HDL region (1.063-1.21 g/ml)
as well as in the 1.21 g/ml infranatant. Examination of the HDL fraction by
electron microscopy revealed round particles, 10-21 nm in diameter. These
data indicate that human apoA-I secreted by transfected 3T3 fibroblasts can
assemble into lipoprotein particles under the appropriate conditions.
Secretion of apolipoprotein A-I in lipoprotein particles following transfection of the human apolipoprotein A-I gene into 3T3 cells
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