title: In vitro factors affecting the concentration of gamma-LpE (gamma-LpE) in human plasma.
abstract: Gamma-LpE (gamma-LpE), a sphingomyelin-rich lipoprotein that contains apolipoprotein (apo) E as its only protein component, has been proposed to play a role in cellular cholesterol efflux by acting, like pre-beta1-LpA-I, as an initial acceptor of cell-derived cholesterol. In order to further characterize the presence of gamma-LpE in human plasma, we have separated gamma-LpE by two-dimensional non-denaturing polyacrylamide-gradient gel electrophoresis and detected its presence by immunoblotting with 125I-labeled polyclonal anti-apoE antibody. Five species of gamma-LpE were routinely detected in human plasma, ranging in mean particle diameter from 9.5 to 16.5 nm. The largest proportion of gamma-migrating apoE was associated with gamma-LpE having a diameter of 13.0 nm. Neither the amount of gamma-LpE apoE (representing less than 1-2% of total plasma apoE) nor the number of gamma-LpE subfractions was different in serum vs. plasma, or was affected by the presence of agents able to inhibit protein dimerization. Gamma-LpE subfractions were present in the plasma of patients having different apoE phenotypes (i.e., apoE 2/2, 3/3, or 4/4). Incubation of plasma at 37 degrees C (90 min) caused a significant decrease in plasma gamma-LpE (>80%) that was not dependent on LCAT or CETP activity. Storage (at -70 degrees C) of hypertriglyceridemic but not normolipidemic plasma resulted in an increase in gamma-LpE. Freezing of postprandial plasma samples, containing increased amounts of triglyceride-rich lipoproteins (TRL) enriched in apoE, also caused an increase in gamma-LpE. Incubation of VLDL (d < 1.006 g/ml) with lipase resulted in the production of gamma-migrating apoE. These results demonstrate that: 1) different gamma-LpE subfractions exist in human plasma; 2) the amount of apoE associated with gamma-LpE subfractions is dependent on in vitro conditions of plasma storage; and 3) TRL can act as a source of gamma-LpE apoE in vitro.
authors.affiliation: Hyperlipidemia and Atherosclerosis Research Group, Clinical Research Institute of Montreal, Quebec, Canada.
authors.forename: L
authors.initials: L
authors.lastname: Krimbou
authors.forename: M
authors.initials: M
authors.lastname: Tremblay
authors.forename: H
authors.initials: H
authors.lastname: Jacques
authors.forename: J
authors.initials: J
authors.lastname: Davignon
authors.forename: J S
authors.initials: JS
authors.lastname: Cohn
mesh_terms: D000328:Adult; D001057:Apolipoproteins E / Q000097:blood*; D016477:Artifacts; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000097:blood; D053480:Cholesterol Ester Transfer Proteins; D002850:Chromatography, Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006023:Glycoproteins; D006801:Humans; D015228:Hypertriglyceridemia / Q000097:blood; D008055:Lipids / Q000097:blood; D008075:Lipoproteins, HDL / Q000097:blood*; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000037:antagonists & inhibitors / Q000097:blood; D013048:Specimen Handling
chemical_list: D001057:Apolipoproteins E; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D006023:Glycoproteins; D008055:Lipids; D008075:Lipoproteins, HDL; D007862:Phosphatidylcholine-Sterol O-Acyltransferase