ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid
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 d...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-07-01
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| Series: | Biology |
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| Online Access: | https://www.mdpi.com/2079-7737/8/3/53 |
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| author | Edward B. Neufeld Masaki Sato Scott M. Gordon Vinay Durbhakula Nicolas Francone Angel Aponte Gizem Yilmaz Denis Sviridov Maureen Sampson Jingrong Tang Milton Pryor Alan T. Remaley |
| author_facet | Edward B. Neufeld Masaki Sato Scott M. Gordon Vinay Durbhakula Nicolas Francone Angel Aponte Gizem Yilmaz Denis Sviridov Maureen Sampson Jingrong Tang Milton Pryor Alan T. Remaley |
| author_sort | Edward B. Neufeld |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T07:56:51Z |
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| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-12T07:56:51Z |
| publishDate | 2019-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-6436e268c8c74166bdf86e2198998d042023-09-02T20:10:46ZengMDPI AGBiology2079-77372019-07-01835310.3390/biology8030053biology8030053ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent PhospholipidEdward B. Neufeld0Masaki Sato1Scott M. Gordon2Vinay Durbhakula3Nicolas Francone4Angel Aponte5Gizem Yilmaz6Denis Sviridov7Maureen Sampson8Jingrong Tang9Milton Pryor10Alan T. Remaley11Lipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USAProteomics Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USADepartment of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALipoprotein Metabolism Laboratory, Translational Vascular Medicine Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USAWe 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.https://www.mdpi.com/2079-7737/8/3/53phospholipids/phosphatidylethanolaminecholesterol/effluxapolipoproteinsHDL (High-density lipoprotein)/metabolismlipoproteinsmass spectrometrymembranes/modellow-density lipoprotein (LDL) |
| spellingShingle | Edward B. Neufeld Masaki Sato Scott M. Gordon Vinay Durbhakula Nicolas Francone Angel Aponte Gizem Yilmaz Denis Sviridov Maureen Sampson Jingrong Tang Milton Pryor Alan T. Remaley ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid Biology phospholipids/phosphatidylethanolamine cholesterol/efflux apolipoproteins HDL (High-density lipoprotein)/metabolism lipoproteins mass spectrometry membranes/model low-density lipoprotein (LDL) |
| title | ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid |
| title_full | ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid |
| title_fullStr | ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid |
| title_full_unstemmed | ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid |
| title_short | ApoA-I-Mediated Lipoprotein Remodeling Monitored with a Fluorescent Phospholipid |
| title_sort | apoa i mediated lipoprotein remodeling monitored with a fluorescent phospholipid |
| topic | phospholipids/phosphatidylethanolamine cholesterol/efflux apolipoproteins HDL (High-density lipoprotein)/metabolism lipoproteins mass spectrometry membranes/model low-density lipoprotein (LDL) |
| url | https://www.mdpi.com/2079-7737/8/3/53 |
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