Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins

Abstract High-density lipoprotein (HDL) particles have multiple beneficial and cardioprotective roles, yet our understanding of their full structural and functional repertoire is limited due to challenges in separating HDL particles from contaminating plasma proteins and other lipid-carrying particl...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jack Jingyuan Zheng, Joanne K. Agus, Brian V. Hong, Xinyu Tang, Christopher H. Rhodes, Hannah E. Houts, Chenghao Zhu, Jea Woo Kang, Maurice Wong, Yixuan Xie, Carlito B. Lebrilla, Emily Mallick, Kenneth W. Witwer, Angela M. Zivkovic
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4becad93e295478eaddaaab1020fc365
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:4becad93e295478eaddaaab1020fc365
record_format	dspace
spelling	oai:doaj.org-article:4becad93e295478eaddaaab1020fc3652021-12-02T16:28:00ZIsolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins10.1038/s41598-021-95451-32045-2322https://doaj.org/article/4becad93e295478eaddaaab1020fc3652021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95451-3https://doaj.org/toc/2045-2322Abstract High-density lipoprotein (HDL) particles have multiple beneficial and cardioprotective roles, yet our understanding of their full structural and functional repertoire is limited due to challenges in separating HDL particles from contaminating plasma proteins and other lipid-carrying particles that overlap HDL in size and/or density. Here we describe a method for isolating HDL particles using a combination of sequential flotation density ultracentrifugation and fast protein liquid chromatography with a size exclusion column. Purity was visualized by polyacrylamide gel electrophoresis and verified by proteomics, while size and structural integrity were confirmed by transmission electron microscopy. This HDL isolation method can be used to isolate a high yield of purified HDL from a low starting plasma volume for functional analyses. This method also enables investigators to select their specific HDL fraction of interest: from the least inclusive but highest purity HDL fraction eluting in the middle of the HDL peak, to pooling all of the fractions to capture the breadth of HDL particles in the original plasma sample. We show that certain proteins such as lecithin cholesterol acyltransferase (LCAT), phospholipid transfer protein (PLTP), and clusterin (CLUS) are enriched in large HDL particles whereas proteins such as alpha-2HS-glycoprotein (A2HSG), alpha-1 antitrypsin (A1AT), and vitamin D binding protein (VDBP) are enriched or found exclusively in small HDL particles.Jack Jingyuan ZhengJoanne K. AgusBrian V. HongXinyu TangChristopher H. RhodesHannah E. HoutsChenghao ZhuJea Woo KangMaurice WongYixuan XieCarlito B. LebrillaEmily MallickKenneth W. WitwerAngela M. ZivkovicNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jack Jingyuan Zheng Joanne K. Agus Brian V. Hong Xinyu Tang Christopher H. Rhodes Hannah E. Houts Chenghao Zhu Jea Woo Kang Maurice Wong Yixuan Xie Carlito B. Lebrilla Emily Mallick Kenneth W. Witwer Angela M. Zivkovic Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
description	Abstract High-density lipoprotein (HDL) particles have multiple beneficial and cardioprotective roles, yet our understanding of their full structural and functional repertoire is limited due to challenges in separating HDL particles from contaminating plasma proteins and other lipid-carrying particles that overlap HDL in size and/or density. Here we describe a method for isolating HDL particles using a combination of sequential flotation density ultracentrifugation and fast protein liquid chromatography with a size exclusion column. Purity was visualized by polyacrylamide gel electrophoresis and verified by proteomics, while size and structural integrity were confirmed by transmission electron microscopy. This HDL isolation method can be used to isolate a high yield of purified HDL from a low starting plasma volume for functional analyses. This method also enables investigators to select their specific HDL fraction of interest: from the least inclusive but highest purity HDL fraction eluting in the middle of the HDL peak, to pooling all of the fractions to capture the breadth of HDL particles in the original plasma sample. We show that certain proteins such as lecithin cholesterol acyltransferase (LCAT), phospholipid transfer protein (PLTP), and clusterin (CLUS) are enriched in large HDL particles whereas proteins such as alpha-2HS-glycoprotein (A2HSG), alpha-1 antitrypsin (A1AT), and vitamin D binding protein (VDBP) are enriched or found exclusively in small HDL particles.
format	article
author	Jack Jingyuan Zheng Joanne K. Agus Brian V. Hong Xinyu Tang Christopher H. Rhodes Hannah E. Houts Chenghao Zhu Jea Woo Kang Maurice Wong Yixuan Xie Carlito B. Lebrilla Emily Mallick Kenneth W. Witwer Angela M. Zivkovic
author_facet	Jack Jingyuan Zheng Joanne K. Agus Brian V. Hong Xinyu Tang Christopher H. Rhodes Hannah E. Houts Chenghao Zhu Jea Woo Kang Maurice Wong Yixuan Xie Carlito B. Lebrilla Emily Mallick Kenneth W. Witwer Angela M. Zivkovic
author_sort	Jack Jingyuan Zheng
title	Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
title_short	Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
title_full	Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
title_fullStr	Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
title_full_unstemmed	Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins
title_sort	isolation of hdl by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of hdl-associated proteins
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4becad93e295478eaddaaab1020fc365
work_keys_str_mv	AT jackjingyuanzheng isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT joannekagus isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT brianvhong isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT xinyutang isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT christopherhrhodes isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT hannahehouts isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT chenghaozhu isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT jeawookang isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT mauricewong isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT yixuanxie isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT carlitoblebrilla isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT emilymallick isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT kennethwwitwer isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins AT angelamzivkovic isolationofhdlbysequentialflotationultracentrifugationfollowedbysizeexclusionchromatographyrevealssizebasedenrichmentofhdlassociatedproteins
_version_	1718383928759287808

Isolation of HDL by sequential flotation ultracentrifugation followed by size exclusion chromatography reveals size-based enrichment of HDL-associated proteins

Ejemplares similares