Both apolipoprotein (apo) E and apoA-I are associated with lipoproteins, although with different particle classes. ApoE is associated with very low density lipoprotein (VLDL) and with the larger high density lipoprotein (HDL) subspecies, while apoA-I is found predominantly in association with most HDL subclasses. The genes encoding these proteins have a similar overall structure with the nucleotide sequences of the third and fourth exons coding for the mature protein. In an effort to understand the difference in lipoprotein association patterns of these two apoproteins, we have constructed and expressed chimeric apoproteins using cDNAs in which the third (n) and fourth (c) exons of human apoE and apoA-I are exchanged. McArdle rat hepatoma cells (McA-RH7777), which secrete VLDL- and HDL-like particles, were stably transfected with these cDNAs, and the cDNAs for human apoE and human apoA-I. Single spin NaBr gradient fractions of lipoprotein deficient serum-treated cell medium from transfected McA-RH7777 cells were analyzed. The distributions of transfected human apoE and apoA-I and endogenous rat apoE and apoA-I were compared with those of the chimeras. Among HDL subspecies, human apoE expressed by these cells is associated with particles of density 1.108 g/ml. Similarly, chimera apoA-InEc (exon 3 of apoA-I and exon 4 of apoE) is found in particles of density 1.111 g/ml. Human apoA-I, however, distributes in a broader range of particles with peak densities of 1.111 g/ml and 1.164 g/ml. The distribution of the complementary chimera, apoEnA-Ic, follows this same pattern, with peak particle densities of 1.098 and 1.137 g/ml. This is in contrast to the narrow distributions of endogenous rat apoE and apoA-I, which were found in particles of density 1.099 and 1.089 g/ml, respectively. When metabolically labeled medium was fractionated via gel filtration column chromatography, apoA-InEc was found to associate with the VLDL fractions; apoEnA-Ic was absent from these same fractions. These results suggest that the fourth exon largely determines the distinctive lipoprotein distribution patterns of these two human apoproteins and that the human apoA-I fourth exon sequence may account for the polydisperse HDL pattern as observed by others in transgenic mice expressing human apoA-I.

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