Association of human apolipoprotein E with lipoproteins secreted by transfected McA RH7777 cells.
To examine the association of apolipoprotein (apo) E with nascent hepatic lipoproteins we have prepared stable transfectants of the rat hepatoma cell line McA RH7777 expressing the human apoE3 cDNA. When the nascent lipoproteins secreted from control cells were separated on fast protein liquid chromatography (FPLC) columns, rat apoE was detected in the very low density (VLDL) and high density lipoprotein (HDL) fractions, while rat apoA-I was found in the HDL and lipoprotein free fractions. Human apoE was also associated with the VLDL and HDL particles secreted from the transfected McA RH7777 cells. Expression of human apoE resulted in a significant decrease in the amount of rat apoA-I associated with the lipoprotein particles. Rat apoE was also displaced, but to a lesser extent. Infection of McA RH7777 cells at different multiplicities of infection with recombinant adenoviral vector containing the human apoE cDNA indicated that rat apoA-I was decreased in the HDL fractions at lower levels of expression of human apoE than was rat apoE. The HDL particles were further examined by immunoblotting of nondenaturing gradient gels and by non-denaturing immunoprecipitation. The results indicate that the high density lipoprotein (HDL) particles are heterogeneous in size and apolipoprotein composition with the majority of the rat and human apolipoproteins being located on different particles. These results suggest that the profile and concentration of HDL apolipoproteins produced in hepatocytes influences the assembly of the various subsets of secreted HDL.
Department of Pathology, The University of Chicago, IL 60637-1470, USA.
C A
CA
Reardon
L
L
Blachowicz
K M
KM
Watson
E
E
Barr
G S
GS
Getz
D000818:Animals; D016632:Apolipoprotein A-I / Q000096:biosynthesis / Q000302:isolation & purification; D001057:Apolipoproteins E / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008075:Lipoproteins, HDL / Q000096:biosynthesis* / Q000302:isolation & purification; D008079:Lipoproteins, VLDL / Q000096:biosynthesis* / Q000302:isolation & purification; D008114:Liver Neoplasms, Experimental; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection; D014407:Tumor Cells, Cultured
D016632:Apolipoprotein A-I; D001057:Apolipoproteins E; D008075:Lipoproteins, HDL; D008079:Lipoproteins, VLDL; D011994:Recombinant Proteins