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Originally published In Press as doi:10.1194/jlr.M300059-JLR200 on June 16, 2003

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Journal of Lipid Research, Vol. 44, 1698-1704, September 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology

PLTP secreted by HepG2 cells resembles the high-activity PLTP form in human plasma

Sarah Siggins, Matti Jauhiainen, Vesa M. Olkkonen, Jukka Tenhunen and Christian Ehnholm1

National Public Health Institute, Department of Molecular Medicine, Biomedicum, P.O. Box 104, FIN-00251 Helsinki, Finland

1 To whom correspondence should be addressed. e-mail: christian.ehnholm{at}ktl.fi

Plasma phospholipid transfer protein (PLTP) is an important regulator of plasma HDL levels and HDL particle distribution. PLTP is present in plasma in two forms, one with high and the other with low phospholipid transfer activity. We have used the human hepatoma cell line, HepG2, as a model to study PLTP secreted from hepatic cells. PLTP activity was secreted by the cells into serum-free culture medium as a function of time. However, modification of a previously established ELISA assay to include a denaturing sample pretreatment with the anionic detergent sodium dodecyl sulphate was required for the detection of the secreted PLTP protein. The HepG2 PLTP could be enriched by Heparin-Sepharose affinity chromatography and eluted in size-exclusion chromatography at a position corresponding to the size of 160 kDa. PLTP coeluted with apolipoprotein E (apoE) but not with apoB-100 or apoA-I. A portion of PLTP was retained by an anti-apoE immunoaffinity column together with apoE, suggesting an interaction between these two proteins. Furthermore, antibodies against apoE but not those against apoB-100 or apoA-I were capable of inhibiting PLTP activity.

These results show that the HepG2-derived PLTP resembles in several aspects the high-activity form of PLTP found in human plasma.

Abbreviations: apoA-I, apolipoprotein A-I; HRP, horseradish peroxidase; IgG, immunoglobulin G

Supplementary key words phospholipid transfer protein • apolipoprotein E • enzyme-linked immunosorbent assay • high density lipoprotein metabolism


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