Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform

Abstract The extracellular vesicle (EV) surface proteome (surfaceome) acts as a fundamental signalling gateway by bridging intra‐ and extracellular signalling networks, dictates EVs’ capacity to communicate and interact with their environment, and is a source of potential disease biomarkers and ther...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Alin Rai, Haoyun Fang, Bethany Claridge, Richard J. Simpson, David W Greening
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	extracellular vesicles mass spectrometry‐based proteomics surface proteins surfaceome vesicle heterogeneity Cytology QH573-671
Acceso en línea:	https://doaj.org/article/0cf32a4a52624a688cb04bd23aca0176
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:0cf32a4a52624a688cb04bd23aca0176
record_format	dspace
spelling	oai:doaj.org-article:0cf32a4a52624a688cb04bd23aca01762021-11-24T14:04:31ZProteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform2001-307810.1002/jev2.12164https://doaj.org/article/0cf32a4a52624a688cb04bd23aca01762021-11-01T00:00:00Zhttps://doi.org/10.1002/jev2.12164https://doaj.org/toc/2001-3078Abstract The extracellular vesicle (EV) surface proteome (surfaceome) acts as a fundamental signalling gateway by bridging intra‐ and extracellular signalling networks, dictates EVs’ capacity to communicate and interact with their environment, and is a source of potential disease biomarkers and therapeutic targets. However, our understanding of surface protein composition of large EVs (L‐EVs, 100–800 nm, mean 310 nm, ATP5F1A, ATP5F1B, DHX9, GOT2, HSPA5, HSPD1, MDH2, STOML2), a major EV‐subtype that are distinct from small EVs (S‐EVs, 30–150 nm, mean 110 nm, CD44, CD63, CD81, CD82, CD9, PDCD6IP, SDCBP, TSG101) remains limited. Using a membrane impermeant derivative of biotin to capture surface proteins coupled to mass spectrometry analysis, we show that out of 4143 proteins identified in density‐gradient purified L‐EVs (1.07–1.11 g/mL, from multiple cancer cell lines), 961 proteins are surface accessible. The surface molecular diversity of L‐EVs include (i) bona fide plasma membrane anchored proteins (cluster of differentiation, transporters, receptors and GPI anchored proteins implicated in cell‐cell and cell‐ECM interactions); and (ii) membrane surface‐associated proteins (that are released by divalent ion chelator EDTA) implicated in actin cytoskeleton regulation, junction organization, glycolysis and platelet activation. Ligand‐receptor analysis of L‐EV surfaceome (e.g., ITGAV/ITGB1) uncovered interactome spanning 172 experimentally verified cognate binding partners (e.g., ANGPTL3, PLG, and VTN) with highest tissue enrichment for liver. Assessment of biotin inaccessible L‐EV proteome revealed enrichment for proteins belonging to COPI/II‐coated ER/Golgi‐derived vesicles and mitochondria. Additionally, despite common surface proteins identified in L‐EVs and S‐EVs, our data reveals surfaceome heterogeneity between the two EV‐subtype. Collectively, our study provides critical insights into diverse proteins operating at the interactive platform of L‐EVs and molecular leads for future studies seeking to decipher L‐EV heterogeneity and function.Alin RaiHaoyun FangBethany ClaridgeRichard J. SimpsonDavid W GreeningTaylor & Francis Grouparticleextracellular vesiclesmass spectrometry‐based proteomicssurface proteinssurfaceomevesicle heterogeneityCytologyQH573-671ENJournal of Extracellular Vesicles, Vol 10, Iss 13, Pp n/a-n/a (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	extracellular vesicles mass spectrometry‐based proteomics surface proteins surfaceome vesicle heterogeneity Cytology QH573-671
spellingShingle	extracellular vesicles mass spectrometry‐based proteomics surface proteins surfaceome vesicle heterogeneity Cytology QH573-671 Alin Rai Haoyun Fang Bethany Claridge Richard J. Simpson David W Greening Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
description	Abstract The extracellular vesicle (EV) surface proteome (surfaceome) acts as a fundamental signalling gateway by bridging intra‐ and extracellular signalling networks, dictates EVs’ capacity to communicate and interact with their environment, and is a source of potential disease biomarkers and therapeutic targets. However, our understanding of surface protein composition of large EVs (L‐EVs, 100–800 nm, mean 310 nm, ATP5F1A, ATP5F1B, DHX9, GOT2, HSPA5, HSPD1, MDH2, STOML2), a major EV‐subtype that are distinct from small EVs (S‐EVs, 30–150 nm, mean 110 nm, CD44, CD63, CD81, CD82, CD9, PDCD6IP, SDCBP, TSG101) remains limited. Using a membrane impermeant derivative of biotin to capture surface proteins coupled to mass spectrometry analysis, we show that out of 4143 proteins identified in density‐gradient purified L‐EVs (1.07–1.11 g/mL, from multiple cancer cell lines), 961 proteins are surface accessible. The surface molecular diversity of L‐EVs include (i) bona fide plasma membrane anchored proteins (cluster of differentiation, transporters, receptors and GPI anchored proteins implicated in cell‐cell and cell‐ECM interactions); and (ii) membrane surface‐associated proteins (that are released by divalent ion chelator EDTA) implicated in actin cytoskeleton regulation, junction organization, glycolysis and platelet activation. Ligand‐receptor analysis of L‐EV surfaceome (e.g., ITGAV/ITGB1) uncovered interactome spanning 172 experimentally verified cognate binding partners (e.g., ANGPTL3, PLG, and VTN) with highest tissue enrichment for liver. Assessment of biotin inaccessible L‐EV proteome revealed enrichment for proteins belonging to COPI/II‐coated ER/Golgi‐derived vesicles and mitochondria. Additionally, despite common surface proteins identified in L‐EVs and S‐EVs, our data reveals surfaceome heterogeneity between the two EV‐subtype. Collectively, our study provides critical insights into diverse proteins operating at the interactive platform of L‐EVs and molecular leads for future studies seeking to decipher L‐EV heterogeneity and function.
format	article
author	Alin Rai Haoyun Fang Bethany Claridge Richard J. Simpson David W Greening
author_facet	Alin Rai Haoyun Fang Bethany Claridge Richard J. Simpson David W Greening
author_sort	Alin Rai
title	Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
title_short	Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
title_full	Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
title_fullStr	Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
title_full_unstemmed	Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
title_sort	proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/0cf32a4a52624a688cb04bd23aca0176
work_keys_str_mv	AT alinrai proteomicdissectionoflargeextracellularvesiclesurfaceomeunravelsinteractivesurfaceplatform AT haoyunfang proteomicdissectionoflargeextracellularvesiclesurfaceomeunravelsinteractivesurfaceplatform AT bethanyclaridge proteomicdissectionoflargeextracellularvesiclesurfaceomeunravelsinteractivesurfaceplatform AT richardjsimpson proteomicdissectionoflargeextracellularvesiclesurfaceomeunravelsinteractivesurfaceplatform AT davidwgreening proteomicdissectionoflargeextracellularvesiclesurfaceomeunravelsinteractivesurfaceplatform
_version_	1718415030617112576

Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform

Ejemplares similares