Efflux of cholesterol from different cellular pools.
Free cholesterol is very efficiently removed from cells by 2-hydroxypropyl-beta-cyclodextrins. The efflux of cholesterol occurs from two distinct kinetic pools: the half-times (t(1/2)) for the two pools in CHO-K1 cells are 15 +/- 5 s and 21 +/- 6 min and they represent 25% +/- 5% and 75% +/- 5% of the readily exchangeable cell cholesterol, respectively. In this study we have determined that the fast pool and the majority of the slow kinetic pool for cholesterol efflux are apparently present in the plasma membrane. Numerous agents that inhibit intracellular cholesterol trafficking are unable to affect either the size or the t(1/2) for efflux of either kinetic pool. In contrast, treatment of the cells with N-ethylmaleimide (NEM), exogenous lipases such as sphingomyelinase and phospholipase C, calcium ionophore A23187, or heat resulted in the dramatic increase in the size of the fast kinetic pool of cholesterol. These changes in the kinetics of cholesterol efflux are not specific to the nature of the extracellular acceptor indicating that they are a consequence of changes in the cell plasma membrane. The above treatments disrupt the normal organization of the lipids in the plasma membrane via either hydrolysis or randomization. The phosphatidylcholine and sphingomyelin present in the plasma membrane are critical for maintaining the two kinetic pools of cholesterol; any alteration in the amount or the location of these phospholipids results in an enhancement of efflux by redistributing cholesterol into the fast kinetic pool.
Haynes
M P
MP
Department of Biochemistry, MCP-Hahnemann University, Philadelphia, Pennsylvania 19129, USA.
Phillips
M C
MC
Rothblat
G H
GH
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D016466:CHO Cells; D000001:Calcimycin / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D002794:Choline / Q000378:metabolism; D006224:Cricetinae; D003505:Cyclodextrins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006358:Hot Temperature; D006868:Hydrolysis / Q000187:drug effects; D007476:Ionophores / Q000494:pharmacology; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism / Q000494:pharmacology; D047392:beta-Cyclodextrins
D003505:Cyclodextrins; D004791:Enzyme Inhibitors; D007476:Ionophores; D008081:Liposomes; D010743:Phospholipids; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000001:Calcimycin; D000255:Adenosine Triphosphate; D002784:Cholesterol; D010738:Type C Phospholipases; D013108:Sphingomyelin Phosphodiesterase; C105598:phosphatidylcholine-specific phospholipase C; D002794:Choline; D005033:Ethylmaleimide