Role of the carboxy-terminal domain of human apolipoprotein AI in high-density-lipoprotein metabolism--a study based on deletion and substitution variants in transgenic mice.

Cholesterol levels in high-density lipoprotein (HDL) of transgenic mice overexpressing human apolipoprotein AI (apoAI), a des-(190-243)-apoAI deletion mutant or an apoAI-(1-189)-apoAII-(12-77) chimera were 2.8-fold (P<0.001), 1.3-fold (P<0.05) and 2.2-fold (P<0.001) higher than in control mice, respectively. Human apolipoprotein levels in apoAI and in apoAI-(1-189)-apoAII-(12-77) transgenic mice were 5.2-fold and 3.5-fold higher than in des-(190-243)-apoAI transgenic mice, whereas their HDL cholesterol levels were 2.1-fold and 1.6-fold higher. PAGE of HDL isolated by ultracentrifugation revealed that murine HDL migrated as 9.6-nm and 7.2-nm particles. Overexpression of human apoAI and apoAI-(1-189)-apoAII-(12-77) resulted in the production of polydisperse HDL (9.6, 9.2, 8.4 and 7.2 nm) particles, whereas overexpression of des-(190-243)-apoAI primarily resulted in an increase of 7.2-nm particles. The fractional catabolic rates of human apoAI and apoAI-(1-189)-apoAII-(12-77) were very similar, whereas that of des-(190-243)-apoAI was 4.9-fold higher. The endogenous production rates of human apoAI, des-(190-243)-apoAI and apoAI-(1-189)-apoAII-(12-77) in transgenic mice were very similar. It is concluded that deletion of the carboxy-terminal domain of apoAI reduces its lipoprotein association, resulting in the production of small, phospholipid-rich HDL particles that are cleared more rapidly. Substitution of the carboxy-terminal helices of apoAI with helices of apoAII restores lipoprotein association, resulting in the production of HDL, which migrates as human HDL3 and HDL2. Although the carboxy-terminal domain of the chimera contained more than 80% of the amino acid sequence of apoAII, its HDL-distribution profile in transgenic mice was very similar to that of human apoAI. This study demonstrates the importance of the helical structure of apoAI of the carboxy-terminal domain of apoAI, rather than of its exclusive amino acid sequence, in HDL metabolism.