Next Article in Journal
Chronotype and Social Jetlag: A (Self-) Critical Review
Previous Article in Journal
The Use of Myelinating Cultures as a Screen of Glycomolecules for CNS Repair
Open AccessArticle

ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid

1
Lipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
2
Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD 20892, USA
3
Proteomics Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
*
Author to whom correspondence should be addressed.
Biology 2019, 8(3), 53; https://doi.org/10.3390/biology8030053
Received: 30 May 2019 / Revised: 1 July 2019 / Accepted: 8 July 2019 / Published: 12 July 2019
(This article belongs to the Section Biochemistry and Molecular Biology)
We describe simple, sensitive and robust methods to monitor lipoprotein remodeling and cholesterol and apolipoprotein exchange, using fluorescent Lissamine Rhodamine B head-group tagged phosphatidylethanolamine (*PE) as a lipoprotein reference marker. Fluorescent Bodipy cholesterol (*Chol) and *PE directly incorporated into whole plasma lipoproteins in proportion to lipoprotein cholesterol and phospholipid mass, respectively. *Chol, but not *PE, passively exchanged between isolated plasma lipoproteins. Fluorescent apoA-I (*apoA-I) specifically bound to high-density lipoprotein (HDL) and remodeled *PE- and *Chol-labeled synthetic lipoprotein-X multilamellar vesicles (MLV) into a pre-β HDL-like particle containing *PE, *Chol, and *apoA-I. Fluorescent MLV-derived *PE specifically incorporated into plasma HDL, whereas MLV-derived *Chol incorporation into plasma lipoproteins was similar to direct *Chol incorporation, consistent with apoA-I-mediated remodeling of fluorescent MLV to HDL with concomitant exchange of *Chol between lipoproteins. Based on these findings, we developed a model system to study lipid transfer by depositing fluorescent *PE and *Chol-labeled on calcium silicate hydrate crystals, forming dense lipid-coated donor particles that are readily separated from acceptor lipoprotein particles by low-speed centrifugation. Transfer of *PE from donor particles to mouse plasma lipoproteins was shown to be HDL-specific and apoA-I-dependent. Transfer of donor particle *PE and *Chol to HDL in whole human plasma was highly correlated. Taken together, these studies suggest that cell-free *PE efflux monitors apoA-I functionality. View Full-Text
Keywords: phospholipids/phosphatidylethanolamine; cholesterol/efflux; apolipoproteins; HDL (High-density lipoprotein)/metabolism; lipoproteins; mass spectrometry; membranes/model; low-density lipoprotein (LDL) phospholipids/phosphatidylethanolamine; cholesterol/efflux; apolipoproteins; HDL (High-density lipoprotein)/metabolism; lipoproteins; mass spectrometry; membranes/model; low-density lipoprotein (LDL)
Show Figures

Figure 1

MDPI and ACS Style

Neufeld, E.B.; Sato, M.; Gordon, S.M.; Durbhakula, V.; Francone, N.; Aponte, A.; Yilmaz, G.; Sviridov, D.; Sampson, M.; Tang, J.; Pryor, M.; Remaley, A.T. ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid. Biology 2019, 8, 53.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop