Protein biocargo of citrus fruit-derived vesicles reveals heterogeneous transport and extracellular vesicle populations

Pocsfalvi, Gabriella; Turiák, Lilla; Ambrosone, Alfredo; Gaudio, Pasquale Del; Puska, Gina; Fiume, Immacolata; Silvestre, Teresa; Vékey, Károly

doi:10.1016/j.jplph.2018.07.006

Cited by 104 publications

(114 citation statements)

References 40 publications

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“…Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the two vesicle fractions purified in the 1 M and 2 M sucrose/D 2 O cushions ( Figure 1B) shows different profiles. The 1 M fraction reveals a highly complex pattern as it is generally observed in EV studies [13]. The yields determined after density cushion UC in the 1 M EV-like fraction are 1.16 × 10 12 particles/L of fruit juice and 1.67 mg of NV proteins/L of fruit juice.…”

Section: Characterization Of Clementine Nanovesiclesmentioning

confidence: 71%

“…[1] shows the numbers of unique and common Orthologous Groups (OGs) of the identified protein. OGs of Citrus clementina (azure) were compared to four citrus species (C. sinensis, C. limon, C. paradise and C. aurantium) (yellow) [13] and EVpedia (red) [14].…”

Section: Isolation Of Nanovesicles (Nvs) From Clementine Juicementioning

confidence: 99%

“…A comparative study was performed to highlight similarities and differences between protein biocargo of clementina vesicles and existing datasets. Orthologous groups (OGs) of each identified protein (678 hits, in Table S2) predicted by EggNOG mapper [15] were searched against (i) the OG accession codes published in EVpedia (27,517 hits, in Table S2) [14] and (ii) four different citrus species (995 unique hits, in Table S2) published recently [13]. The Venn diagram in Figure 1C shows the high overlap percentages found with both EVpedia (548 hits, 85%) and four citrus data sets (543 hit, 84%).…”

Section: The Protein Cargo Of Citrus Clementina Nanovesiclesmentioning

confidence: 99%

“…Fruit juices are one of the most popular sources to isolate edible plant-derived vesicles, as they are readily available by squeezing. By studying the protein biocargo of citrus fruit-derived vesicles, Pocsfalvi et al have demonstrated the presence of highly expressed HSP70, HSP80, 14-3-3, G3PD and FBA6, PTL3 and clathirin proteins along with a large number of different enzymes and membrane transporters [13].…”

Section: Introductionmentioning

confidence: 99%

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Membrane Transporters in Citrus clementina Fruit Juice-Derived Nanovesicles

Stanly

Moubarak

Fiume

et al. 2019

IJMS

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The cellular vesicle is a fluid-filled structure separated from the surrounding environment by a biological membrane. Here, we isolated nanovesicles (NVs) from the juice of clementines using a discontinuous density gradient ultracentrifugation method. To gain information about the protein content of vesicles, mass spectrometry-based organelle proteomics and bioinformatics were applied to the exosome-like vesicle fraction isolated in the 1 mol/L sucrose/D2O cushion. Analysis of 1018 identified proteins revealed a highly complex mixture of different intra, extracellular and artificially-formed vesicle populations. In particular, clathrin-coated vesicles were significantly expressed in this sample. Membrane transporters are significantly represented in clementines nanovesicles. We have found 162 proteins associated with the transport Gene Ontology term (GO: 0006810) which includes; 71 transmembrane transport related, 53 vesicle mediated and 50 intracellular transporters. Platellin-3 like carrier protein containing a Sec14 domain is known to have a role in plant-virus interaction and that is one of the most abundant proteins in our dataset. The presence of transmembrane transporters like ATPases, aquaporins, ATP Binding Cassette (ABC) transporters and tetraspanins, regulators of protein trafficking suggests that nanovesicles of clementines can actively interact with their environment in a controlled way.

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Section: Characterization Of Clementine Nanovesiclesmentioning

confidence: 71%

Section: Isolation Of Nanovesicles (Nvs) From Clementine Juicementioning

confidence: 99%

Section: The Protein Cargo Of Citrus Clementina Nanovesiclesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Membrane Transporters in Citrus clementina Fruit Juice-Derived Nanovesicles

Stanly

Moubarak

Fiume

et al. 2019

IJMS

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“…These issues motivated researchers to explore alternative EV sources, in particular, food such as bovine milk and plants, which have the main advantage of being easily accessible, cost‐effective, and scalable . Since they are common components of our diet, milk and plants are also considered biocompatible and safe sources of vesicles .…”

Section: Methods For Improvement Of Upstream Ev Yieldmentioning

confidence: 99%

Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Paganini

Palmiero

Pócsfalvi

et al. 2019

Biotechnology Journal

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Potential applications of extracellular vesicles (EVs) are attracting increasing interest in the fields of medicine, cosmetics, and nutrition. However, the manufacturing of EVs is currently characterized by low yields. This limitation severely hampers progress in research at the laboratory and clinical scales, as well as the realization of successful and cost-effective EV-based products. Moreover, the high level of heterogeneity of EVs further complicates reproducible manufacturing on a large scale. In this review, possible directions toward the scalable production of EVs are discussed. In particular, two strategies are considered: i) the optimization of upstream unit operations and ii) the exploitation of well-established and mature technologies already in use in other industrial bioprocesses.

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Plant‐Derived Vesicle‐Like Nanoparticles as Promising Biotherapeutic Tools: Present and Future

et al. 2023

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At present, it is generally acknowledged that EVs are secreted by almost all living cells including plant cells. [4] Compared with mammalian EVs, the research of plant EVs is still in its infancy. Increasing evidence has proved that plant cells are able to secret exosome-like vesicles or extracellular vesicles into apoplast space. [5] In the 1960s, using transmission electron microscopy (TEM), Halperin et al. for the first time observed and discovered multivesicular bodies (MVBs) fusing with the plasma membrane and releasing exosome-like vesicles into the cell wall area in carrot cell cultures. [6] Fifty years after this first discovery, Regenteet al. used a tissue-infiltration-centrifugation method and successfully separated exosome-like vesicles from apoplast washing fluids (AWF) of sunflower seeds. [5a] Following these discoveries, plant exosomelike vesicles or vesicle-like nanoparticles have been successfully isolated from many plant species, including ginseng, ginger, broccoli, bitter melon, oat, grape, grapefruit, lemon, blueberry, orange, sunflower seed, and Arabidopsis, etc. [5a,7] The efficient separation of PDVLNs from plant cells is still one of the challenges that need to be solved in the plant EV field. At present, two plant tissue pre-processing methods, namely tissue-disruption and tissue-infiltration centrifugation, have emerged as favorite methods used for vesicle separation from plant cells in the field. [5b,8] After tissue pre-treatment, most studies use ultracentrifugation or density gradient centrifugation for further PDVLN separation. [5a,9] In addition, novel separation methods are under development for PDVLNs, such as ultrafiltration, immunoaffinity capture, size-exclusive chromatography, etc. [5b,7j,10] However, it is still challenging to isolate specific PDVLN subtypes, due to the lack of generic or specific markers for PDVLNs and well-verified plant species antibodies. [4b,5b] The nomenclature of vesicles prepared by these methods is not standardized in existing references. Many terms were used, including "plant particles", "plant vesicles", etc. [7b,8b,10,11] To facilitate communication in the plant vesicle field, standardizing the nomenclatures is necessary. We thus propose to unify these nomenclatures into plant-derived vesicle-like nanoparticles (PDVLNs) in this review.PDVLNs share many similar physical characteristics to those of human-derived EVs, including features such as morphology, particle size, concentration, Zeta potential, etc. [2a,7b,e,11] To further elucidate the specific molecular mechanism of PDVLNs on Extracellular vesicles (EVs) are heterogeneous, phospholipid bilayer-enclosed biological particles that regulate cell communication by molecular cargo delivery and surface signaling. EVs are secreted by almost all living cells, including plant cells. Plant-derived vesicle-like nanoparticles (PDVLNs) is a generic term referring to vesicle-like nanostructure particles isolated from plants. Their low immunogenicity and wide availability make PDVLNs safer and mor...

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Protein biocargo of citrus fruit-derived vesicles reveals heterogeneous transport and extracellular vesicle populations

Cited by 104 publications

References 40 publications

Membrane Transporters in Citrus clementina Fruit Juice-Derived Nanovesicles

Membrane Transporters in Citrus clementina Fruit Juice-Derived Nanovesicles

Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Plant‐Derived Vesicle‐Like Nanoparticles as Promising Biotherapeutic Tools: Present and Future

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