Proteomic and Biochemical Analyses of Human B Cell-derived Exosomes

By R. Wubbolts, R. Leckie, P. Veenhuizen, G. Schwarzmann, W. Möbius, Joerg Hoernschemeyer, J. Slot, H. Geuze, W. Stoorvogel. Journal of Biological Chemistry, 2003

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Exosomes are 60–100-nm membrane vesicles that are secreted into the extracellular milieu as a consequence of multivesicular body fusion with the plasma membrane. Here we determined the protein and lipid compositions of highly purified human B cell-derived exosomes. Mass spectrometric analysis indicated the abundant presence of major histocompatibility complex (MHC) class I and class II, heat shock cognate 70, heat shock protein 90, integrin α4, CD45, moesin, tubulin (α and β), actin, Giα2, and a multitude of other proteins. An α4-integrin may direct B cell-derived exosomes to follicular dendritic cells, which were described previously as potential target cells. Clathrin, heat shock cognate 70, and heat shock protein 90 may be involved in protein sorting at multivesicular bodies. Exosomes were also enriched in cholesterol, sphingomyelin, and ganglioside GM3, lipids that are typically enriched in detergent-resistant membranes. Most exosome-associated proteins, including MHC class II and tetraspanins, were insoluble in 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS)-containing buffers. Multivesicular body-linked MHC class II was also resistant to CHAPS whereas plasma membrane-associated MHC class II was solubilized readily. Together, these data suggest that recruitment of membrane proteins from the limiting membranes into the internal vesicles of multivesicular bodies may involve their incorporation into tetraspanin-containing detergent-resistant membrane domains.

Subfields: Immunology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/6f68b7f1836c90c22bf36b144fbe2295030a3699

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Citations in the corpus

Extracellular vesicles: Exosomes, microvesicles, and friends (2013, background): “In comparison to the plasma membrane, exosomes from a variety of cells (Wubbolts et al., 2003; Laulagnier et al., 2004; Subra et al., 2007; Brouwers et al., 2012) are highly enriched in cholesterol, sphingomyelin, and cells grown in fetal calf serum depleted of EVs by performing differential…”
Introduction to Extracellular Vesicles: Biogenesis, RNA Cargo Selection, Content, Release, and Uptake (2016, neutral): “Adding to this observation is the abundance of ceramide and its derivatives in exosomes, as well as the presence of proteolipoprotein (PLP), CD63, CD81, and TSG101 (Wubbolts et al. 2003; Brouwers et al. 2013).”
Exosome Function: From Tumor Immunology to Pathogen Biology (2008, background): “Similar to proteins, lipids present on exosomes are characteristic of the cell origin, with most of the lipid analytical work being performed on exosomes derived from reticulocytes (33), mast cells (34), B lymphocyte cell lines (35) and human DCs (34).”
Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. (2012, background): “other sphingolipids, and phospoglycerides with long and saturated fatty-acyl chains [12,18,19].”
Exosomes--vesicular carriers for intercellular communication. (2009, background): “However, such vesicles do not appear to play a role in exosome formation, as mass spectrometry analysis of the lipid composition of exosomes from human B-cells revealed that BMP/LBPA is absent from exosomes [12].”
Functional proteomic screens reveal an essential extracellular role for hsp90α in cancer cell invasiveness (2004, background): “One possibility is that hsp90α might be released through exosomes from tumour cells, as observed with B cell”
Exosomal-like vesicles are present in human blood plasma. (2005, methodology): “The lipid composition of exosomes reveals an enrichment in cholesterol, sphingomyelin and gangliosid GM3 levels ( 24 , 27).”
Cell-Produced α-Synuclein Is Secreted in a Calcium-Dependent Manner by Exosomes and Impacts Neuronal Survival (2010, result): “In agreement with previous studies (Wubbolts et al., 2003; Valadi et al., 2007; Simpson et al., 2008; Gonzalez-Begne et al., 2009), LC-MS proteomic analysis of our exosome preparation identified common, as well as cell-type-specific, proteins (supplemental table, available at www.”
Review of the Isolation, Characterization, Biological Function, and Multifarious Therapeutic Approaches of Exosomes (2019, background): “Exosomes originating from a variety of different cells share common structural and functional proteins, such as Rab GTPase, SNAREs, annexins, Alix, Tsg101, and tetraspanins [13,14] and the glycosylphosphatidylinositol-anchored molecules, flotillin, cholesterol, sphingomyelin, and hexosylceramides [1”
Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies. (2014, background): “ILVs accumulate cholesterol during their biogenesis from the endosome (MVB) membrane, and subsequently exosomes are enriched in cholesterol as compared to the parental cells [37,83].”
Mesenchymal stem cell: an efficient mass producer of exosomes for drug delivery. (2013, background): “Their membranes are enriched in lipid rafts, cholesterol, sphingomyelin and ceramide [21,22].”
Classification, Functions, and Clinical Relevance of Extracellular Vesicles (2012, background): “…exosomes are surrounded by a phospholipid membrane containing relatively high levels of cholesterol, sphingomyelin, and ceramide and containing detergent-resistant membrane domains (lipid rafts) (Wubbolts et al., 2003; Simons and Raposo, 2009; Théry et al., 2009; Mathivanan et al., 2010).”