J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on August 1, 2005

Papers In Press, published online ahead of print May 16, 2005
J. Lipid Res., doi:10.1194/jlr.M500038-JLR200
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Submitted on February 1, 2005
Revised on April 12, 2005
Accepted on May 2, 2005

Biogenesis and speciation of nascent apoA-I-containing particles in various cell lines

Larbi Krimbou, Houssein Hajj Hassan, Sacha Blain, Shirya Rashid, Maxime Denis, Michel Marcil, and Jacques Genest

Department of Cardiology, McGill University Health Center, Montreal, QC H3A 1A1

Corresponding Author: jacques.genest{at}muhc.mcgill.ca

It is generally thought that the large heterogeneity of human HDL confers antiatherogenic properties, however, the mechanisms governing HDL biogenesis and speciation are complex and poorly understood. Here, we show that incubation of exogenous apoA-I with either fibroblasts, CaCo-2 or CHO-overexpressing ABCA1 cells generates only alpha LpA-I with diameters of 8 to 20 nm, whereas HUVEC and ABCA1 mutant (Q597R) cells were unable to form such particles. Interestingly, incubation of exogenous apoA-I with either HepG2 or macrophages generates both alpha LpA-I and prebeta 1LpA-I. Furthermore, glyburide inhibits almost completely the formation of alpha LpA-I but not prebeta 1LpA-I. Similarly, endogenously secreted HepG2 apoA-I was found associated with both prebeta 1LpA-I and alpha LpA-I, by contrast, CaCo-2 cells secreted only alpha LpA-I. To determine whether alpha LpA-I generated by fibroblasts is a good substrate for LCAT, isolated alpha LpA-I as well as rHDL was reacted with LCAT. Although both particles had similar Vmax (8.4 vs. 8.2 nmol CE/h/µg LCAT, respectively), the Km value was increased 2-fold for alpha LpA-I compared to rHDL (1.2 vs. 0.7 µM apoA-I). These results demonstrate that: 1) ABCA1 is required for the formation of alpha LpA-I but not prebeta 1LpA-I; and 2) nascent alpha LpA-I interacts efficiently with LCAT. Thus, our study provides direct evidence for a new link between specific cell lines and the speciation of nascent HDL that occurs by both ABCA1-dependent and -independent pathways.


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