Proteomic dissection of large extracellular vesicle surfaceome unravels interactive surface platform
journal contributionposted on 2021-12-15, 23:34 authored by A Rai, H Fang, Bethany ClaridgeBethany Claridge, Richard SimpsonRichard Simpson, David GreeningDavid Greening
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.
National Health and Medical Research Council, Grant/Award Numbers: 1057741, 1139489; Future Fund, Grant/Award Number: 1201805; Helen Amelia Hains Fellowship; Victorian State Government Operational Infrastructure; Australian Government Training Program; Baker Institute Bright Sparks Scholarship Top Up
JournalJournal of Extracellular Vesicles
Article NumberARTN e12164
Rights StatementThe Author reserves all moral rights over the deposited text and must be credited if any re-use occurs. Documents deposited in OPAL are the Open Access versions of outputs published elsewhere. Changes resulting from the publishing process may therefore not be reflected in this document. The final published version may be obtained via the publisher’s DOI. Please note that additional copyright and access restrictions may apply to the published version.
Science & TechnologyLife Sciences & BiomedicineCell Biologyextracellular vesiclesmass spectrometry-based proteomicssurface proteinssurfaceomevesicle heterogeneityPRECURSOR-LIKE PROTEIN-2CELL-LINE LIM1863GLYCOLYTIC-ENZYMESCANCER CELLSINTERCELLULAR TRANSFERDRUG-RESISTANCECOLON-CARCINOMAQUANTITATIVE PROTEOMICSP-GLYCOPROTEINEXOSOMES