Methylamine-treated low density lipoproteins elicit different responses in HepG2 cells and macrophages.

Abstract

Recent results from this laboratory have demonstrated the existence of labile thiolester bonds in apolipoprotein B (ApoB). Thiolester bonds can be cleaved with nucleophiles such as methylamine, resulting in conformational change. The purpose of this study was to explore whether the cellular interactions would be altered after methylamine treatment of low density lipoproteins (LDL). Human hepatoma cells, HepG2, and human monocyte derived macrophages were used for these studies. Fresh LDL were incubated with methylamine under mild alkaline conditions under N2 and with preservatives for 24 h. The methylamine-treated LDL showed particle size and net charge identical to fresh native LDL. In addition, no oxidative modification of LDL occurred under the experimental conditions. The methylamine-treated LDL were indistinguishable from native LDL in HepG2 cells as judged by binding, degradation, cholesterol accumulation and de novo sterol synthesis. However, methylamine-treated LDL caused an increased accumulation of cholesteryl esters in macrophages which was comparable to the accumulation caused by acetylated LDL. Dual color digital imaging fluorescence microscopy revealed no competition between acetylated and methylamine-treated LDL, suggesting that the excessive uptake of methylamine-treated LDL was not mediated by the 'scavenger' receptor. The increased accumulation of cholesteryl ester in macrophages also did not appear to stem from the classical LDL receptor. These results suggest that a new receptor binding domain is exposed due to the conformational change upon treatment of LDL with methylamine.