title: Interfacial and lipid transfer properties of human phospholipid transfer protein: implications for the transfer mechanism of phospholipids.
abstract: In circulation the phospholipid transfer protein (PLTP) facilitates the transfer of phospholipid-rich surface components from postlipolytic chylomicrons and very low density lipoproteins (VLDL) to HDL and thereby regulates plasma HDL levels. To study the molecular mechanisms involved in PLTP-mediated lipid transfer, we studied the interfacial properties of PLTP using Langmuir phospholipid monolayers and asymmetrical flow field-flow fractionation (AsFlFFF) to follow the transfer of 14C-labeled phospholipids and [35S]PLTP between lipid vesicles and HDL particles. The AsFlFFF method was also used to determine the sizes of spherical and discoidal HDL particles and small unilamellar lipid vesicles. In Langmuir monolayer studies high-activity (HA) and low-activity (LA) forms of PLTP associated with fluid phosphatidylcholine monolayers spread at the air/buffer interphase. Both forms also mediated desorption of [14C]dipalmitoylphosphatidylcholine (DPPC) from the phospholipid monolayer into the buffer phase, even when it contained no physiological acceptor such as HDL. After the addition of HDL3 to the buffer, HA-PLTP caused enhanced lipid transfer to them. The particle diameter of HA-PLTP was approximately 6 nm and that of HDL3 approximately 8 nm as determined by AsFlFFF analysis. Using this method, it could be demonstrated that in the presence of HA-PLTP, but not LA-PLTP, [14C]DPPC was transferred from small unilamellar vesicles (SUV) to acceptor HDL3 molecules. Concomitantly, [35S]-HA-PLTP was transferred from the donor to acceptor, and this transfer was not observed for its low-activity counterpart. These observations suggest that HA-PLTP is capable of transferring lipids by a shuttle mechanism and that formation of a ternary complex between PLTP, acceptor, and donor particles is not necessary for phospholipid transfer.
authors.affiliation: Department of Ophthalmology, University of Helsinki, Helsinki, Finland.
authors.forename: Niko L
authors.initials: NL
authors.lastname: Setälä
authors.forename: Juha M
authors.initials: JM
authors.lastname: Holopainen
authors.forename: Jari
authors.initials: J
authors.lastname: Metso
authors.forename: Susanne K
authors.initials: SK
authors.lastname: Wiedmer
authors.forename: Gebrenegus
authors.initials: G
authors.lastname: Yohannes
authors.forename: Paavo K J
authors.initials: PK
authors.lastname: Kinnunen
authors.forename: Christian
authors.initials: C
authors.lastname: Ehnholm
authors.forename: Matti
authors.initials: M
authors.lastname: Jauhiainen
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism; D002250:Carbon Radioisotopes; D006801:Humans; D050356:Lipid Metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D010316:Particle Size; D049294:Phospholipid Transfer Proteins / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D013462:Sulfur Radioisotopes
chemical_list: D002250:Carbon Radioisotopes; D008075:Lipoproteins, HDL; D008081:Liposomes; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; D013462:Sulfur Radioisotopes; D015060:1,2-Dipalmitoylphosphatidylcholine