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Green fluorescent protein‐tagged apolipoprotein E: A useful marker for the study of hepatic lipoprotein egress
Author(s) -
Takacs Constantin N.,
Andreo Ursula,
Belote Rachel L.,
Pulupa Joan,
Scull Margaret A.,
Gleason Caroline E.,
Rice Charles M.,
Simon Sanford M.
Publication year - 2017
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12467
Subject(s) - apolipoprotein e , biology , lipoprotein , secretion , green fluorescent protein , microbiology and biotechnology , apolipoprotein b , lipid anchored protein , biogenesis , hepatocyte , biochemistry , cholesterol , medicine , disease , gene , autophagy , in vitro , apoptosis
Apolipoprotein E ( ApoE ), a component of very‐low‐density and high‐density lipoproteins, participates in many aspects of lipid transport in the bloodstream. Underscoring its important functions, ApoE isoforms have been associated with metabolic and circulatory disease. ApoE is also incorporated into hepatitis C virus ( HCV ) particles, and promotes their production and infectivity. Live cell imaging analysis of ApoE behavior during secretion from producing cells thus has the potential to reveal important details regarding lipoprotein and HCV particle biogenesis and secretion from cells. However, this approach requires expression of fluorescently tagged ApoE constructs that need to faithfully reproduce known ApoE behaviors. Herein, we evaluate the usefulness of using an ApoE‐GFP fusion protein in studying hepatocyte‐derived, ApoE ‐containing lipoproteins and HCV particles. We show that while ApoE‐GFP alone is not sufficient to support infectious HCV production, it nonetheless colocalizes intracellularly and associates with secreted untagged lipoprotein components. Furthermore, its rate of secretion from hepatic cells is indistinguishable from that of untagged ApoE . ApoE‐GFP thus represents a useful marker for ApoE ‐containing hepatic lipoproteins.