Insect lipid transfer particle catalyzes bidirectional vectorial transfer of diacylglycerol from lipophorin to human low density lipoprotein.
Insect plasma lipid transfer particle (LTP) catalyzes vectorial net transfer of diacylglycerol (DAG) from Manduca sexta larval high density lipophorin (HDLp-L) to human low density lipoprotein (LDL) producing an LDL of lower density and lipophorin subspecies of higher density. At equilibrium, a stable DAG-depleted very high density lipophorin species (density = 1.25 g/ml) is formed. Electrophoretic analysis of the substrate and product lipoproteins showed that apoprotein exchange or transfer between human LDL and lipophorin did not occur during the lipid transfer reaction. Facilitated net transfer of cholesteryl ester, free cholesterol, and phospholipid occurred to a much lower extent than DAG net transfer, indicating that under these conditions, LDL serves as a sink for lipophorin-associated DAG. This reaction, therefore, provides a method whereby the mass of lipid associated with human LDL can be modified in vitro without alteration of its apoprotein component. The DAG content of LDL increased in a linear manner with respect to LTP concentration and time during the initial phase of the reaction, demonstrating the utility of this system as a quantitative assay method for LTP-mediated net DAG transfer. When [3H]DAG-labeled LDL was prepared and employed in transfer experiments with unlabeled lipophorin, labeled DAG was recovered in the HDLp-L fraction. The amount of labeled DAG recovered in the HDLp-L fraction was dependent on the ratio of LDL to HDLp-L in the reaction. Thus, in this system, LTP-mediated DAG redistribution is bidirectional, suggesting that the final equilibrium distribution of lipid may be dictated by the properties of potential donor/acceptor lipoproteins rather than by an inherent particle substrate specificity of LTP.
Ryan
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Lipid and Lipoprotein Research Group, University of Alberta, Edmonton, Canada.
Wessler
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Price
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Ando
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Yokoyama
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D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005989:Glycerides / Q000378:metabolism*; D006458:Hemolymph / Q000378:metabolism; D006801:Humans; D007814:Larva; D007915:Lepidoptera / Q000378:metabolism*; D008074:Lipoproteins / Q000302:isolation & purification; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008970:Molecular Weight; D009036:Moths / Q000378:metabolism*; D011485:Protein Binding
D002352:Carrier Proteins; D004075:Diglycerides; D005989:Glycerides; D008074:Lipoproteins; D008077:Lipoproteins, LDL; C030363:lipid transfer protein; C032830:lipophorin