EV-TRACK

Search > Results

You searched for: EV180055 (EV-TRACK ID)

Showing 1 - 1 of 1

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV180055	1/1	Leishmania infantum	Leishmania infantum	(d)(U)C Filtration	Santarém N	2012	42%
Study summary Full title Exoproteome dynamics in Leishmania infantum. All authors Santarém N, Racine G, Silvestre R, Cordeiro-da-Silva A, Ouellette M Journal Proteomics Abstract The exoproteome of Leishmania infantum is composed of parasite derived proteins present in the extra (show more...)The exoproteome of Leishmania infantum is composed of parasite derived proteins present in the extracellular environment. Although the exoproteome might have a significant role in the precocious steps of infection little is known concerning its composition. We developed an approach enabling the in vitro recovery of the exoproteome from logarithmic and stationary L. infantum promastigotes. The recovered exoproteomes were further separated into two fractions, vesicles and vesicle depleted exoproteome, evaluating the fraction protein profile. Although the most abundant protein in all fractions was GP63, the protein composition of the separated fractions was distinct reflecting the origin of the fraction and the metabolic state of the parasites. The vesicle-derived exoproteome recovered from logarithmic parasites was significantly enriched in ribosomal proteins, indicating a potential role for these vesicles in protein turnover. Also, a stage specific enrichment of vesicles with properties related to apoptotic vesicles was observed in stationary phase parasites and evidence was obtained that the release of vesicles was increased in response to a death stimuli. This report on the exoproteome obtained from in vitro promastigote cultures provides new perspectives on Leishmania biology with the possibility of vesicles playing a major role in protein turnover and also in cell death. BIOLOGICAL SIGNIFICANCE: The first systematic insight into Leishmania exoproteome composition and the impact of the selected recovery approach. (hide) EV-METRIC 42% (80th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C Filtration Adj. k-factor 255.8 (pelleting) Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Function Sample Species Leishmania infantum Sample Type Cell culture supernatant EV-producing cells Leishmania infantum EV-harvesting Medium Serum free medium Cell viability (%) NA Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180 Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 100000 Pelleting: adjusted k-factor 255.8 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 30-100 EM EM-type Transmission-EM Image type Wide-field Extra information The reported work is related to the characterization of extracelular vesicles from protozoan parasites, Leishmania infantum, using diferente culture conditions to produce the vesicles. In the same publication is also analysed vesicle depleated fractions. The recovery strategy reflects the need to recover both vesicles and also the vesicle depleated fraction from the same preparation.

				1 - 1 of 1

EV-TRACK ID	EV180055
species	Leishmania infantum
sample type	Cell culture
cell type	Leishmania infantum
condition	Control condition
separation protocol	(d)(U)C Filtration
Exp. nr.	1
EV-METRIC %	42