J. Lipid Res.
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A more recent version of this article appeared on November 1, 2002

Papers In Press, published online ahead of print September 1, 2002
J. Lipid Res., doi:10.1194/jlr.D200024-JLR200
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Submitted on June 28, 2002
Revised on September 1, 2002
Accepted on August 20, 2002

The differential apolipoprotein A-I enrichment of preb1 and alpha-HDL is detectable by gel filtration separation

Maud Chetiveaux, Hassan Nazih, Veronique Ferchaud-Roucher, Gilles Lambert, Yassine zair, Martial Masson, Khadija Ouguerram, Daniele Bouhours, and Michel Krempf

Inserm U539, Nantes, Cedex 01 44093

Corresponding Author: maud.chetiveaux{at}sante.univ-nantes.fr

The aim of the study was to assess the isolation of HDL by Fast Protein Liquid Chromatography (FPLC) to perform kinetics studies of Apo A-I-HDL labelled with a stable isotope. Comparison between FPLC and ultracentrifugation have been made. Apo A-I-HDL kinetics were studied by infusion of [5.5.5-2H3]-leucine for 14 hours in five subjects. Using FPLC, preb1 HDL and aHDL (HDL2 and HDL3) were separated from 200µl of plasma samples. Total HDL were isolated by sequential ultracentrifugation (HDL-UC). The tracer-to-tracee ratio were higher in preb1 HDL than in total HDL-UC. The higher leucine enrichment found in total HDL-UC compared to aHDL suggested the existence of a mixture of Apo A-I-HDL sub-classes. From this difference in enrichments, the turnover rate of total HDL-UC, usually assumed to be aHDL was probably overestimated in previous studies. To our knowledge, this study is the first report which provides a convenient tool to distinguish enrichments of Apo A-I in preb1 HDL and aHDL from total HDL previously used for kinetic measurements. This original and new method should help to understand the kinetics of HDL in humans and the reverse cholesterol transport dynamics.


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