Domains of apoE4 required for the biogenesis of apoE-containing HDL

Abstract

Introduction. We have studied the functions of truncated apoE4 forms in vitro and in vivo in order to identify the domains of apoE4 required for the biogenesis of apoE-containing high-density lipoprotein (HDL). Results. We have found that apoE4-185, -202, -229, or -259 could promote ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux in vitro, although less efficiently than Full-length apoE4, and had diminished capacity to activate lecithin cholesterol acyltransferase (LCAT). Formation of HDL in vivo was assessed by various methods following gene transfer in apolipoprotein A-I^−/− × apoE^−/− mice. Fast protein liquid chromatography of plasma showed that the truncated apoE forms, except apoE4-185, generated an apoE-containing HDL peak. Two-dimensional gel electrophoresis of plasma and electron microscopy showed that truncated apoE forms generated distinct HDL subpopulations and formed discoidal HDL particles which could be converted to spherical by co-administration of truncated apoE4-202 and LCAT. Conclusion. Overall, the in-vivo and in-vitro data are consistent and indicate that apoE4-185 is the shortest truncated form that supports formation of discoidal apoE4-containing HDL particles.

Key words::

• Apolipoprotein E
• HDL biogenesis
• HDL gene transfer
• LCAT

Acknowledgements

We thank Konstantinos Drosatos, Andreas Kateifides, Gayle Forbes, and Adelina Duka for technical assistance and Anne Plunkett for preparing the manuscript.

Declaration of interest: This work was supported by grants from the National Institutes of Health (HL68216 and HL48739). A Vezeridis was supported by the NIH Predoctoral Training Grant (HL007969).