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Candida albicans (ATCC90028 and ATCC10231 yeast) extracellular vesicle and whole cell lysate proteomes

ABSTRACT: We performed proteomic analysis of extracellular vesicles (EVs) and whole cell lysates (WCL) isolated from the same liquid culture. This was implemented for two different C. albicans strains, ATCC90028 (n = 3) and ATCC10231 (n = 3). This data set is the first label-free quantitative comparative study of the proteins in C. albicans EVs and their parent planktonic (yeast-form) cells. We identified proteins enriched in EVs versus WCL by comparing their LFQ intensities across all samples. After linking these results with EV enrichment data from other strains, we proposed a suite of putative EV marker proteins which were useful for multiple C. albicans strains and morphologies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Candida Albicans (yeast)

SUBMITTER: Charlotte Dawson

LAB HEAD: Mark Bleackley

PROVIDER: PXD014388 | Pride | 2020-04-17

REPOSITORIES: Pride

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			Action	DRS
	10231_EV1.raw	Raw
	10231_EV1_20190212233737.raw	Raw
	10231_EV2.raw	Raw
	10231_EV2_20190213061715.raw	Raw
	10231_EV3.raw	Raw

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Publications

Protein markers for <i>Candida albicans</i> EVs include claudin-like Sur7 family proteins.

Dawson Charlotte S CS Garcia-Ceron Donovan D Rajapaksha Harinda H Faou Pierre P Bleackley Mark R MR Anderson Marilyn A MA

Journal of extracellular vesicles 20200416 1

<b>Background:</b> Fungal extracellular vesicles (EVs) have been implicated in host-pathogen and pathogen-pathogen communication in some fungal diseases. In depth research into fungal EVs has been hindered by the lack of specific protein markers such as those found in mammalian EVs that have enabled sophisticated isolation and analysis techniques. Despite their role in fungal EV biogenesis, ESCRT proteins such as Vps23 (Tsg101) and Bro1 (ALIX) are not present as fungal EV cargo. Furthermore, tet ...[more]

PMID: 32363014

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Candida albicans (DAY286 yeast) extracellular vesicle and whole cell lysate proteomes

Project description:We performed proteomic analysis of extracellular vesicles (EVs) and whole cell lysates (WCL) isolated from the same liquid culture (strain = DAY286, n = 3). This data set is the first label-free quantitative comparative study of the proteins in C. albicans EVs and their parent planktonic (yeast-form) cells. We identified proteins enriched in EVs versus WCL by comparing their LFQ intensities across all samples. After linking these results with EV enrichment data from other strains, we proposed a suite of putative EV marker proteins which were useful for multiple C. albicans strains and morphologies.

2020-04-17 | PXD014367 | Pride

Candida albicans (DAY286 biofilm) extracellular vesicle and whole cell lysate proteomes

Project description:We performed proteomic analysis of extracellular vesicles (EVs) and whole cell lysates (WCL) isolated from the same biofilm (strain = DAY286, n = 5). We identified proteins enriched in EVs versus WCL by comparing their LFQ intensities across all samples. After linking these results with EV enrichment data from other strains, we proposed a suite of putative EV marker proteins which were useful for multiple C. albicans strains and morphologies.

2020-04-17 | PXD014389 | Pride

Proteome of extracellular vesicles from the filamentous fungal pathogen Fusarium graminearum

Project description:We performed a comparative study to determine the proteome of extracellular vesicles (EVs) from the cereal pathogen Fusarium graminearum (Fgr), recovered from in vitro cultures. Label-free quantitative proteomics was used to find significant enrichment of proteins between EV samples, the secretome (secreted-soluble proteins) and the cell lysate. Our results show that some proteins were exclusive to EVs and were upregulated compared to the secretome or cell lysate.

2022-02-16 | PXD028657 | Pride

Fusarium oxysporum f. sp. vasinfectum (UP000030701) extracellular vesicle, secretome, and cell lysate proteomes

Project description:We performed a comparative study to determine the proteome of extracellular vesicles (EVs) from the cotton pathogen Fusarium oxysporum f. sp. vasinfectum (Fov), recovered from two growth conditions in vitro. Label-free quantitative protemics was used to find significant enrichment of proteins between EV samples, the secretome (secreted-soluble proteins) and the cell lysate. Our results show that some proteins were exclusive to EVs and were upregulated compared to the secretome or cell lysate.

2021-03-08 | PXD023907 | Pride

Crosstalk between the effects of extracellular vesicles (Evs) and TNF on U937 human monocyte cell line

Project description:Under physiological conditions, extracellular vesicles (EVs) are present simultaneously in the extracellular compartment together with cytokines. Thus, we hypothesized that EVs in combination with cytokines induce different responses of monocyte cells compared to EVs or cytokines alone. Human monocyte U937 cells were incubated with EV-containing or EV-free CCRF human T-cell supernatant, with or without the addition of TNF. U937 cells cultured in EV-free supernatant, supernatant containing CCRF t-cell derived EVs, TNF or both. Each treatment option was measured in 3 replicates.

2013-06-19 | E-GEOD-47897 | biostudies-arrayexpress

Proteomic Analysis of Mycobacterium tuberculosis Protein-Mannosyltransferase Knock-Out Mutant.

Project description:This project focused in the proteomic characterization of the secreted proteome of a Rv1002c (Protein mannosyltransferase) knock-out mutant and its wild type counterpart. The analysis was carried out using a M. tuberculosis ΔleuDΔpanCD auxotroph Rv1002c wild type compared to M. tuberculosis ΔleuDΔpanCDΔRv1002c. Both strains were grown in triplicate and their whole cell lysate (WCL) and secreted proteins (CFP) where purified, trypsin digested and characterized by liquid-chromatography coupled with tandem mass spectrometry (LC-MS/MS). Three replicate injections were performed for each of the sampels. The resulting spectra were searched against an M. tuberculosis database and relative protein abundance was determined using normalized spectral counts. Protein abundance differences were tested by Student’s t test. Abundance of 39 secreted proteins was significantly different between the WT and the ΔRv1002c. Decrease abundance of known glycoproteins, including several lipoglycoproteins was observed in the ΔRv1002c strain. In the WCL, 61 proteins presented significantly different abundance between the WT and the ΔRv1002c strains, including 4 proteins that were completely absent in the mutant (Rv2510c, PknF, Mpt70 and CeoC).

2019-04-09 | PXD011081 | Pride

Coordination of fungal biofilm development by extracellular vesicle cargo

Project description:The fungal pathogen Candida albicans can form biofilms that protect it from drugs and the immune system. The biofilm cells release extracellular vesicles (EVs) that promote extracellular matrix formation and resistance to antifungal drugs. Here, we define functions for numerous EV cargo proteins in biofilm matrix assembly and drug resistance, as well as in fungal cell adhesion and dissemination. We use a machine-learning analysis of cargo proteomic data from mutants with EV production defects to identify 63 candidate gene products for which we construct mutant and complemented strains for study. Among these, 17 mutants display reduced biofilm matrix accumulation and antifungal drug resistance. An additional subset of 8 cargo mutants exhibit defects in adhesion and/or dispersion. Representative cargo proteins are shown to function as EV cargo through the ability of exogenous wild-type EVs to complement mutant phenotypic defects. Most functionally assigned cargo proteins have roles in two or more of the biofilm phases. Our results support that EVs provide community coordination throughout biofilm development in C. albicans.

2022-02-16 | PXD028596 | Pride

Osteoblasts secrete miRNA-containing extracellular vesicles that enhance expansion of human umbilical cord blood cells

Project description:Osteolineage cells represent one of the critical bone marrow niche components that support maintenance of hematopoietic stem and progenitor cells (HSPCs). Recent studies demonstrate that extracellular vesicles (EVs) regulate stem cell development via horizontal transfer of bioactive cargo, including microRNAs (miRNAs). Here, we characterize the miRNA profile of EVs secreted by human osteoblasts and study their biological effect of on human umbilical cord blood-derived CD34+ HSPCs by sequencing, gene expression and biochemical analyses. Using next-generation sequencing we show that osteoblast-derived EVs contain highly abundant miRNAs specifically enriched in EVs, including critical regulators of hematopoietic proliferation (e.g., miR-29a). EV treatment of CD34+ HSPCs alters the expression of candidate miRNA targets, such as HBP1, BCL2 and PTEN. Furthermore, EVs enhance proliferation of CD34+ cells and their immature subsets in growth factor-driven ex vivo expansion cultures. Importantly, EV-expanded cells retain their differentiation capacity in vitro and show successful engraftment in NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice in vivo. These discoveries reveal a novel osteoblast-derived EV-mediated mechanism for regulation of HSPC proliferation and warrant consideration of EV-miRNAs for the development of expansion strategies to treat hematological disorders.

2016-10-05 | E-MTAB-4560 | biostudies-arrayexpress

Human iPSC-derived cardiomyocytes under hypoxia/normoxia, treated with bone marrow or cardiac stromal cell extracellular vesicles

Project description:RNA-seq was performed on cultured human induced pluripotent cell derived cardiomyocytes (iPSC-CMs). There are four groups, with three samples per group: H1,H2,H3. Negative control. Healthy, normoxic iPSC-CMs D1,D2,D3. Positive control. iPSCs cultured under hypoxic (0-1% O2) for 48h with 2% v/v PBS as vehicle control EV1,EV2,EV3. Treatment 1. iPSCs cultured under hypoxic (0-1% O2) for 48h, with cardiac stromal cell derived extracellular vesicles, provided at a dose of 67ng/µl (2% v/v) EV21, EV23, EV24. Treatment 2 iPSCs cultured under hypoxic (0-1% O2) for 48h, with bone marrow mesenchymal stromal cell (BM-MSC) derived extracellular vesicles, provided at a dose of 67ng/µl (2% v/v) All EVs were isolated from cultured human cells using sequential centrifugation methods. Cells were cultured using commercial EV-depleted FBS to avoid contamination of bovine EVs. EVs were validated for CD81, CD9, ALIX positivity, and visualised by cryoEM. Particle to protein ratios were not different between cardiac and bone marrow EV isolates. Therefore EV doses were standardised so that the same dose by protein (67ng/µl) and EV number (~2,000 EVs per iPSC-CM) were added.

2024-10-02 | E-MTAB-13966 | biostudies-arrayexpress

Proteomic signatures of extracellular vesicles secreted by non-mineralizing and mineralizing human osteoblasts and stimulation of tumor cell growth

Project description:Beyond forming bone, osteoblasts play pivotal roles in various biological processes, including hematopoiesis and bone metastasis. Extracellular vesicles (EVs) have recently been implicated in intercellular communication via transfer of proteins and nucleic acids between cells. Here, we focused on the proteomic characterization of non-mineralizing (NMOBs) and mineralizing (MOBs) human osteoblast (SV-HFOs) EVs and investigated their effect on human prostate cancer (PC3) cells by microscopic, proteomic and gene expression analyses. Proteomic analysis showed that 97% of the proteins were shared among NMOB and MOB EVs, and 30% were novel osteoblast-specific EV proteins. Label-free quantification demonstrated mineralization stage-dependent five-fold enrichment of 59 and 451 EV proteins in NMOBs and MOBs, respectively. Interestingly, bioinformatic analyses of the osteoblast EV proteomes and EV-regulated prostate cancer gene expression profiles showed that they converged on pathways involved in cell survival and growth. This was verified by in vitro proliferation assays where osteoblast EV uptake led to two-fold increase in PC3 cell growth compared to cell-free culture medium-derived vesicle controls. Our findings elucidate the mineralization stage-specific protein content of osteoblast-secreted EVs, show a novel way by which osteoblasts communicate with prostate cancer, and open up innovative avenues for therapeutic intervention. PC3 cells were treated with extracellular vesicles from non-mineralizing and mineralizing SV-HFOs for three different incubation times (4hrs, 24hrs, 48hr)

2014-10-28 | E-GEOD-56172 | biostudies-arrayexpress

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