Internalization of phospholipids from the plasma membrane of human osteoblasts depends on the lipid head group

The redistribution of spin- or fluorescence-labeled phospholipid analogs across the plasma membrane of human osteoblast cells, either in suspension or grown as monolayers, was investigated. After incorporation into the outer membrane leaflet, analogs of the aminophospholipids phosphatidylserine and phosphatidylethanolamine moved rapidly to the inner monolayer, whereas the choline-containing analogs of phosphatidylcholine and sphingomyelin disappeared more slowly from the outer leaflet. The fast inward movement of the aminophospholipids became reduced after lowering the intracellular ATP, suggesting the presence of an aminophospholipid translocase activity in the plasma membrane of these cells. From these data, a transverse phospholipid asymmetry in osteoblasts can be inferred with the aminophospholipids mainly concentrated in the inner monolayer and the choline-containing phospholipids in the outer leaflet. A similar pattern of phospholipid internalization was inferred for osteoblasts from human osteoporotic bones and for a human osteosarcoma cell line. The relevance of the enrichment of phosphatidylserine in the cytoplasmic membrane leaflet for calcification in skeletal tissues is emphasized.