Substrate specificity of human plasma phospholipid transfer protein.
The ability of human plasma phospholipid transfer protein to transfer L-alpha-[14C]dipalmitoylphosphatidylcholine (DPPC) from donor vesicles to acceptor high-density lipoproteins (HDL) was examined, using vesicles of different compositions and sizes, and native or chemically modified HDL. Phosphatidylcholine (PC) transfer was inhibited by both cholesterol and sphingomyelin incorporation into egg-PC vesicles. On a molar basis, cholesterol inhibited transfer about 5-fold more than sphingomyelin; however, the effects of both lipids on the fluidity of the vesicle membrane (measured by fluorescence polarization of diphenylhexatriene), were closely correlated with their effects on PC transfer activity. Increase in vesicle size, and decrease in bilayer curvature, also reduced transfer: the largest vesicles had no transfer activity at all. Addition of phosphatidic acid up to 17 mol% had no effect on PC transfer. HDL apolipoprotein lysyl residues were chemically modified by reductive methylation, citraconylation, or acetoacetylation. The effects of modification on the apolipoprotein structure and on the HDL particle were assessed by intrinsic fluorescence measurements, SDS-polyacrylamide gel electrophoresis patterns, and gel chromatography. Only acetoacetylation significantly affected any of these parameters. The ability of HDL to accept PC in the absence of phospholipid transfer protein decreased with an increase in apolipoprotein negative charge while, in the presence of phospholipid transfer protein, the acceptor ability of HDL increased up to 1.7-fold with an initial increase in negative charge and then decreased, ultimately to zero, upon extensive modification.
D000818:Animals; D002250:Carbon Radioisotopes; D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification; D002417:Cattle; D006801:Humans; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000378:metabolism; D008565:Membrane Proteins; D008745:Methylation; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids / Q000097:blood*; D011663:Pulmonary Surfactants / Q000378:metabolism; D013379:Substrate Specificity
D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D008075:Lipoproteins, HDL; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; D011663:Pulmonary Surfactants