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Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions

High-density lipoprotein (HDL) is a mixture of complex particles mediating reverse cholesterol transport (RCT) and several cytoprotective activities. Despite its relevance for human health, many aspects of HDL-mediated lipid trafficking and cellular signaling remain elusive at the molecular level. D...

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Autores principales: Frey, Kathrin, Goetze, Sandra, Rohrer, Lucia, von Eckardstein, Arnold, Wollscheid, Bernd
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9409371/
https://www.ncbi.nlm.nih.gov/pubmed/36012766
http://dx.doi.org/10.3390/ijms23169506
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author Frey, Kathrin
Goetze, Sandra
Rohrer, Lucia
von Eckardstein, Arnold
Wollscheid, Bernd
author_facet Frey, Kathrin
Goetze, Sandra
Rohrer, Lucia
von Eckardstein, Arnold
Wollscheid, Bernd
author_sort Frey, Kathrin
collection PubMed
description High-density lipoprotein (HDL) is a mixture of complex particles mediating reverse cholesterol transport (RCT) and several cytoprotective activities. Despite its relevance for human health, many aspects of HDL-mediated lipid trafficking and cellular signaling remain elusive at the molecular level. During HDL’s journey throughout the body, its functions are mediated through interactions with cell surface receptors on different cell types. To characterize and better understand the functional interplay between HDL particles and tissue, we analyzed the surfaceome-residing receptor neighborhoods with which HDL potentially interacts. We applied a combination of chemoproteomic technologies including automated cell surface capturing (auto-CSC) and HATRIC-based ligand–receptor capturing (HATRIC-LRC) on four different cellular model systems mimicking tissues relevant for RCT. The surfaceome analysis of EA.hy926, HEPG2, foam cells, and human aortic endothelial cells (HAECs) revealed the main currently known HDL receptor scavenger receptor B1 (SCRB1), as well as 155 shared cell surface receptors representing potential HDL interaction candidates. Since vascular endothelial growth factor A (VEGF-A) was recently found as a regulatory factor of transendothelial transport of HDL, we next analyzed the VEGF-modulated surfaceome of HAEC using the auto-CSC technology. VEGF-A treatment led to the remodeling of the surfaceome of HAEC cells, including the previously reported higher surfaceome abundance of SCRB1. In total, 165 additional receptors were found on HAEC upon VEGF-A treatment representing SCRB1 co-regulated receptors potentially involved in HDL function. Using the HATRIC-LRC technology on human endothelial cells, we specifically aimed for the identification of other bona fide (co-)receptors of HDL beyond SCRB1. HATRIC-LRC enabled, next to SCRB1, the identification of the receptor tyrosine-protein kinase Mer (MERTK). Through RNA interference, we revealed its contribution to endothelial HDL binding and uptake. Furthermore, subsequent proximity ligation assays (PLAs) demonstrated the spatial vicinity of MERTK and SCRB1 on the endothelial cell surface. The data shown provide direct evidence for a complex and dynamic HDL receptome and that receptor nanoscale organization may influence binding and uptake of HDL.
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spelling pubmed-94093712022-08-26 Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions Frey, Kathrin Goetze, Sandra Rohrer, Lucia von Eckardstein, Arnold Wollscheid, Bernd Int J Mol Sci Article High-density lipoprotein (HDL) is a mixture of complex particles mediating reverse cholesterol transport (RCT) and several cytoprotective activities. Despite its relevance for human health, many aspects of HDL-mediated lipid trafficking and cellular signaling remain elusive at the molecular level. During HDL’s journey throughout the body, its functions are mediated through interactions with cell surface receptors on different cell types. To characterize and better understand the functional interplay between HDL particles and tissue, we analyzed the surfaceome-residing receptor neighborhoods with which HDL potentially interacts. We applied a combination of chemoproteomic technologies including automated cell surface capturing (auto-CSC) and HATRIC-based ligand–receptor capturing (HATRIC-LRC) on four different cellular model systems mimicking tissues relevant for RCT. The surfaceome analysis of EA.hy926, HEPG2, foam cells, and human aortic endothelial cells (HAECs) revealed the main currently known HDL receptor scavenger receptor B1 (SCRB1), as well as 155 shared cell surface receptors representing potential HDL interaction candidates. Since vascular endothelial growth factor A (VEGF-A) was recently found as a regulatory factor of transendothelial transport of HDL, we next analyzed the VEGF-modulated surfaceome of HAEC using the auto-CSC technology. VEGF-A treatment led to the remodeling of the surfaceome of HAEC cells, including the previously reported higher surfaceome abundance of SCRB1. In total, 165 additional receptors were found on HAEC upon VEGF-A treatment representing SCRB1 co-regulated receptors potentially involved in HDL function. Using the HATRIC-LRC technology on human endothelial cells, we specifically aimed for the identification of other bona fide (co-)receptors of HDL beyond SCRB1. HATRIC-LRC enabled, next to SCRB1, the identification of the receptor tyrosine-protein kinase Mer (MERTK). Through RNA interference, we revealed its contribution to endothelial HDL binding and uptake. Furthermore, subsequent proximity ligation assays (PLAs) demonstrated the spatial vicinity of MERTK and SCRB1 on the endothelial cell surface. The data shown provide direct evidence for a complex and dynamic HDL receptome and that receptor nanoscale organization may influence binding and uptake of HDL. MDPI 2022-08-22 /pmc/articles/PMC9409371/ /pubmed/36012766 http://dx.doi.org/10.3390/ijms23169506 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Frey, Kathrin
Goetze, Sandra
Rohrer, Lucia
von Eckardstein, Arnold
Wollscheid, Bernd
Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title_full Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title_fullStr Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title_full_unstemmed Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title_short Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions
title_sort decoding functional high-density lipoprotein particle surfaceome interactions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9409371/
https://www.ncbi.nlm.nih.gov/pubmed/36012766
http://dx.doi.org/10.3390/ijms23169506
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