Transfer of cholesterol and oxysterol derivatives by the nonspecific lipid transfer protein (sterol carrier protein 2): a study on its mode of action.
The nonspecific lipid transfer protein (nsLTP) facilitates the transfer of both phospholipids and cholesterol between membrane interfaces. In this study, we have investigated the transport of 14C-labelled cholesterol, 7-ketocholesterol, 7 alpha-hydroxycholesterol and 25-hydroxycholesterol from a mixed lipid monolayer at the air/water interface to acceptor vesicles in the subphase. In the absence of nsLTP the transport of cholesterol was virtually nil, whereas the spontaneous transport of the oxysterol derivatives increased in the order 7-ketosterol less than 7 alpha-hydroxycholesterol less than 25-hydroxycholesterol. In the presence of nsLTP, the transport of both cholesterol and the oxysterol derivatives was greatly enhanced; the highest rate of transport was observed for 25-hydroxycholesterol. In the absence of vesicles, binding of cholesterol and of 25-hydroxycholesterol from the monolayer to nsLTP was negligible. Similarly, nsLTP did not bind cholesterol from radiolabeled bovine heart mitochondria under conditions where it stimulated the transfer of cholesterol to vesicles. In agreement with this failure to bind, nsLTP was unable to carry cholesterol between two separate monolayers. From the monolayer experiments it became apparent that nsLTP is highly surface-active. Measurement of the transport of cholesterol and of oxysterol derivatives by the monolayer-vesicles assay and of a series of pyrene-labeled phosphatidylcholine species by the fluorescent transfer assay showed a high correlation between the spontaneous and the nsLTP-mediated lipid transport. This supports the notion that nsLTP lowers the energy barrier for the lipid monomer-membrane interface equilibration process. In view of the above observations, we propose that nsLTP may facilitate the transfer of lipids by being part of a transient collisional complex between donor and acceptor membrane.
van Amerongen
A
A
Center for Biomembranes and Lipid Enzymology, State University of Utrecht, The Netherlands.
Demel
R A
RA
Westerman
J
J
Wirtz
K W
KW
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000502:physiology*; D002417:Cattle; D002784:Cholesterol / Q000378:metabolism*; D066298:In Vitro Techniques; D008567:Membranes, Artificial; D008929:Mitochondria, Heart / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010940:Plant Proteins; D011485:Protein Binding; D013261:Sterols / Q000378:metabolism*
D002352:Carrier Proteins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010940:Plant Proteins; D013261:Sterols; C072087:sterol carrier proteins; D002784:Cholesterol