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Originally published In Press as doi:10.1194/jlr.M500038-JLR200 on May 16, 2005
Journal of Lipid Research, Vol. 46, 1668-1677, August 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology
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 Genest1
Cardiovascular Genetics Laboratory, Cardiology Division, McGill University Health Centre/Royal Victoria Hospital, Montréal, Québec H3A 1A1, Canada
Published, JLR Papers in Press, May 16, 2005. DOI 10.1194/jlr.M500038-JLR200
1 To whom correspondence should be addressed. e-mail: 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 apolipoprotein A-I (apoA-I) with fibroblasts, CaCo-2, or CHO-overexpressing ABCA1 cells generates only -nascent apolipoprotein A-I-containing particles ( -LpA-I) with diameters of 820 nm, whereas human umbilical vein endothelial cells and ABCA1 mutant (Q597R) cells were unable to form such particles. Interestingly, incubation of exogenous apoA-I with either HepG2 or macrophages generates both -LpA-I and preß1-LpA-I. Furthermore, glyburide inhibits almost completely the formation of -LpA-I but not preß1-LpA-I. Similarly, endogenously secreted HepG2 apoA-I was found to be associated with both preß1-LpA-I and -LpA-I; by contrast, CaCo-2 cells secreted only -LpA-I. To determine whether -LpA-I generated by fibroblasts is a good substrate for LCAT, isolated -LpA-I as well as reconstituted HDL [r(HDL)] was reacted with LCAT. Although both particles had similar Vmax (8.4 vs. 8.2 nmol cholesteryl ester/h/µg LCAT, respectively), the Km value was increased 2-fold for -LpA-I compared with r(HDL) (1.2 vs. 0.7 µM apoA-I).
These results demonstrate that 1) ABCA1 is required for the formation of -LpA-I but not preß1-LpA-I; and 2) -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.
Abbreviations: apoA-I, apolipoprotein A-I; CE, cholesteryl ester; HUVEC, human umbilical vein endothelial cells; LpA-I, nascent apolipoprotein A-I-containing particle; MWCO, molecular weight cut off; RCT, reverse cholesterol transport; r(HDL), reconstituted high density lipoprotein; TD, Tangier disease; 2D-PAGGE, two-dimensional polyacrylamide nondenaturing gradient gel electrophoresis; 22OH/9CRA, 22(R)-hydroxycholesterol and 9-cis-retinoic acid Supplementary key words ATP binding cassette transporter A1 high density lipoprotein -nascent apolipoprotein A-I-containing particle pre-ß1-nascent apolipoprotein A-I-containing particle

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Copyright © 2005 by the American Society for Biochemistry and Molecular Biology.
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